Abstract
There is provided an apparatus for use in cell processing comprising: a holding element arranged to receive a top section of a container within a first plane; a moveable plate, spaced apart from the holding element, arranged to operably engage a base section of a container, the moveable plate defining a second plane substantially parallel to the first plane; and an actuation mechanism operably coupled to the moveable plate to rotate the moveable plate about at least one axis within the second plane, thereby reducing a distance between at least a portion of the second plane and the first plane. There is also provided a method of processing cells.
Claims
1. An apparatus for use in performing one or more unit operations in cell processing comprising: a holding element arranged to receive a top section of a container within a first plane; a moveable plate, spaced apart from the holding element, arranged to operably engage a base section of a container, the moveable plate defining a second plane substantially parallel to the first plane; and an actuation mechanism operably coupled to the moveable plate to rotate the moveable plate about at least one axis within the second plane, thereby reducing a distance between at least a portion of the second plane and the first plane.
2. The apparatus according to claim 1, wherein the actuation mechanism is arranged to rotate the moveable plate about an axis within the second plane.
3. The apparatus according to claim 1, wherein the actuation mechanism is arranged to rotate the moveable plate about a plurality of axes within the second plane.
4. The apparatus according to claim 1, wherein the actuation mechanism is arranged to rotate the moveable plate between 0 degrees and 90 degrees about the at least one axis.
5. The apparatus according to claim 1, wherein a longitudinal axis extending perpendicularly to the first plane and the second plane, intersects the second plane at an origin, wherein the actuation mechanism is arranged to pivot the moveable plate about the origin.
6. The apparatus according to claim 5, wherein the origin is centrally located within the second plane.
7. The apparatus according to claim 5, wherein the moveable plate is pivotable about the origin such that each point within the second plane, excluding the origin, forms an angle with respect to the longitudinal axis between 0 and 180 degrees, excluding 90 degrees.
8. The apparatus according to claim 1, wherein the actuation mechanism is further arranged to move the moveable plate along a longitudinal axis, the longitudinal axis substantially perpendicular to the first plane and the second plane, to reduce a distance between the second plane and the first plane.
9. The apparatus according to claim 1, wherein the holding element comprises: a platform operably engageable with a top section of a container, a clamping mechanism operably engageable with a top section of a container, or a sealing plate operably engageable with a top section of a container.
10. The apparatus according to claim 1, wherein the actuation mechanism comprises a base plate spaced apart from, and substantially parallel to, the moveable plate, the base plate operably coupled to the moveable plate.
11. The apparatus according to claim 10, wherein the base plate comprises at least one actuator, the at least one actuator operably coupled to, or operably engageable with, the moveable plate.
12. The apparatus according to claim 10, wherein the base plate comprises at least one rail, the at least one rail upstanding from the base plate substantially perpendicularly to the base plate, the moveable plate slidably coupled to the at least one rail, and wherein at least one actuator is arranged to slide at least a portion of the moveable plate along the at least one rail.
13. The apparatus according to any claim 10, wherein the base plate comprises a first linkage operably coupled to the moveable plate, the first linkage drivable by a first motor.
14. The apparatus according to claim 13, wherein the base plate further comprises a second linkage operably coupled to the moveable plate, the second linkage driven by a second motor.
15. The apparatus according to claim 14, wherein the first linkage and the second linkage are operably coupled at opposing edges of the moveable plate.
16. The apparatus according to claim 14, wherein the base plate further comprises a third linkage operably coupled to the moveable plate, the third linkage driven by a third motor.
17. The apparatus according to claim 16, wherein the first linkage, the second linkage and the third linkage are operably coupled to the moveable plate in a triangular arrangement.
18. The apparatus according to claim 1, wherein the actuation mechanism comprises a first motor, operably coupled to the moveable plate, and configured to rotate the moveable plate about the at least one axis within the second plane.
19. The apparatus according to claim 18, wherein the actuation mechanism further comprises a second motor, operably coupled to the moveable plate, and configured to move the moveable plate along a longitudinal axis, substantially perpendicular to the first plane and the second plane, to reduce a distance between at least a portion of the second plane and the first plane.
20. The apparatus according to claim 19, wherein the actuation mechanism further comprises a third motor, operably coupled to the moveable plate, and configured to move the moveable plate along the longitudinal axis, to reduce a distance between at least a portion of the second plane and the first plane.
21. The apparatus according to claim 20, wherein the actuation mechanism further comprises a linkage, the linkage comprising: a crank housing, comprising a rotatable crank, slidable along a first longitudinal rail, extending substantially parallel to the longitudinal axis; and a slider portion, operably coupled to the moveable plate, slidable along a second longitudinal rail, extending substantially parallel to the first longitudinal rail, the slider portion operably coupled to the rotatable crank by a connecting rod, wherein the second motor is operably coupled to rotate the rotatable crank and move the slider portion along the second longitudinal rail, and wherein the third motor is operably coupled to the crank housing so as to move the crank housing along the first longitudinal rail.
22. The apparatus according to claim 21, wherein the third motor comprises a ball screw motor or a lead screw motor, operably coupled to a corresponding screw threaded portion of the crank housing, wherein the crank housing is moveable along the first longitudinal rail.
23. The apparatus according to claim 21, further comprising a controller communicatively coupled to the actuation mechanism.
24. The apparatus according to claim 23, further comprising one or more positioning sensors, at least one positioning senor communicatively coupled to the controller, wherein the controller configured to generate a signal to the actuation mechanism based upon a signal received from the at least one position sensor.
25. A system for use in performing one or more unit operations in cell processing comprising: an apparatus comprising: a holding element arranged to receive a top section of a container within a first plane, a moveable plate, spaced apart from the holding element, arranged to operably engage a base section of a container, the moveable plate defining a second plane substantially parallel to the first plane; and an actuation mechanism operably coupled to the moveable plate to rotate the moveable plate about at least one axis within the second plane, thereby reducing a distance between at least a portion of the second plane and the first plane; and a container having a base section, a top section in parallel to the base section, and a compressible wall element extending substantially perpendicularly therebetween, wherein the container is at least partially disposed between the holding element and the moveable plate, the actuation mechanism arranged to move the moveable plate such that at least a portion of the compressible wall element is at least partially compressed along a longitudinal axis perpendicular to the first plane and the second plane.
26. The system according to claim 25, wherein the base section of the container is fixedly attached to the moveable plate.
27. A method of processing cells, the method comprising: providing a cell processing medium in a container, the container having a base section, a top section in parallel to the base section, and a compressible wall element extending substantially perpendicularly therebetween; holding the top section of the container within a first plane; engaging the base section of the container with a moveable plate defining a second plane substantially parallel to, and spaced apart from, the first plane; and rotating the base section of the container about at least one axis within the second plane, to cause turbulence of the cell processing medium within the container.
28. The method according to claim 27, wherein the step of rotating the base section of the container comprises rotating the base section of the container about an axis within the second plane.
29. The method according to claim 27, wherein a longitudinal axis, extending perpendicularly to the first plane and the second plane, intersects the second plane at an origin, wherein the step of rotating the base section of the container comprises pivoting the base section of the container about the origin.
30. The method according to claim 29, wherein the origin is centrally located within the second plane.
31. A method of processing cells, the method comprising: (a) providing a compressible container comprising a population of cells in a liquid medium; and (b) maintaining the population of cells in the liquid medium, while pressure is applied to at least a portion of the compressible container to compress the compressible container.
32. The method according to claim 31, wherein the liquid medium is retained in the compressible container after compression is applied.
33. (canceled)
34. (canceled)
35. (canceled)
Description
BRIEF DESCRIPTION OF THE DRAWINGS
[0386] Example embodiments of the disclosure are now described, by way of example only, hereinafter with reference to the accompanying drawings, in which:
[0387] FIGS. 1(a) and 1(b) illustrates an example of the prior art;
[0388] FIGS. 2(a) to 2(d) illustrate an example of the prior art;
[0389] FIG. 3 illustrates (a) a top view, (b) a side view in a first configuration and (c) a side view in a second configuration of the apparatus according to one embodiment of the disclosure;
[0390] FIG. 4 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0391] FIG. 5 illustrates (a) a side view in a first configuration and (b) a side view in a second configuration of the apparatus according to another embodiment of the disclosure;
[0392] FIG. 6 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0393] FIG. 7 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0394] FIG. 8 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0395] FIG. 9 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0396] FIG. 10 illustrates (a) a first side view, (b) a second side view and (c) the first side view having the central hub removed of the apparatus according to another embodiment of the disclosure, and (d) one example of a cam member and (e) another example of a cam member for use in the apparatus of FIGS. 10(a) to 10(c);
[0397] FIG. 11 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0398] FIG. 12 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0399] FIG. 13 illustrates (a) a side view in a first configuration, (b) a side view in a second configuration and (c) an enlarged view of the apparatus according to another embodiment of the disclosure;
[0400] FIG. 14 illustrates (a) a side view in a first configuration and (b) a side view in a second configuration of the apparatus according to another embodiment of the disclosure;
[0401] FIG. 15 illustrates a side view of the apparatus according to another embodiment of the disclosure;
[0402] FIG. 16 illustrates (a) a side view, (b) a perspective view and (c) another perspective view including a container, of the apparatus according to another embodiment of the disclosure;
[0403] FIG. 17 illustrates (a) a side view in a first configuration and (b) a side view in a second configuration of the apparatus according to another embodiment of the disclosure;
[0404] FIG. 18 illustrates (a) a top view, (b) a first side view, (c) a perspective view and (d) another side view of the apparatus according to another embodiment of the disclosure;
[0405] FIG. 19 illustrates a side view a linear actuator for use in the apparatus according to the disclosure;
[0406] FIG. 20 illustrates (a) a container including water and food coloring prior to agitation and (b) the container including water and food coloring after a swirling agitation;
[0407] FIG. 21 illustrates (a) a container including water and food coloring prior to agitation and (b) a container including water and food coloring after a wave agitation;
[0408] FIGS. 22(a) to 22(c) illustrate a container having water and food coloring during a swirling agitation;
[0409] FIGS. 23(a) to 23(c) illustrate a container having water and food coloring during another swirling agitation;
[0410] FIGS. 24(a) to 24(c) illustrate a container having water and food coloring during yet another swirling agitation;
[0411] FIGS. 25(a) and 25(b) illustrate a container having water and food coloring during another swirling agitation;
[0412] FIGS. 26(a) to 26(c) illustrate a container having water and food coloring during a linear compression agitation;
[0413] FIGS. 27(a) to 27(c) illustrate a container having water and food coloring during another linear compression agitation;
[0414] FIGS. 28(a) to 28(c) illustrate a container having water and food coloring during yet another linear compression agitation;
[0415] FIG. 29 illustrates (a) a side view of a container in a first configuration and (b) a side view of a container in a second configuration;
[0416] FIGS. 30(a) to 30(e) illustrate a system, including a container and an apparatus according to the disclosure, throughout the compression motion;
[0417] FIG. 31 illustrates (a) a side view of a system including a container and an apparatus according to the disclosure in a first configuration, (b) a side view of the system in a second configuration after rotation about an axis and (c) a side view of the system in a third configuration;
[0418] FIG. 32 illustrates a mold for a container;
[0419] FIG. 33 illustrates (a) a top down view, (b) a side view, (c) a bottom view and (d) an exploded perspective view of a platform and containers suitable for use with the apparatus and system according to the disclosure;
[0420] FIG. 34 illustrates (a) a perspective view of a schematic of the apparatus according to another embodiment of the disclosure, (b) a front view of the schematic of (a), (c) a schematic view of one motor and linkage of the apparatus of (a), and (d) a schematic view of another motor and linkage of the apparatus of (a);
[0421] FIG. 35 illustrates a detailed perspective view of the apparatus of FIGS. 34(a) and (b);
[0422] FIG. 36 illustrates (a) a front view, (b) a front perspective view, (c) a rear perspective view, (d) a side view and (e) a top view of the apparatus of FIG. 35 having the platform and containers of FIGS. 33(a) to 33(d) attached thereto;
[0423] FIG. 37 illustrates a method according to the disclosure;
[0424] FIG. 38 illustrates an exemplary testing run timeline;
[0425] FIG. 39 illustrates the results of an exemplary primary T-cell run timeline using the apparatus/system according to the disclosure;
[0426] FIG. 40 illustrates the results of another exemplary primary T-cell run timeline using the apparatus/system according to the disclosure;
[0427] FIG. 41 illustrates the further results from the exemplary primary T-cell run timeline illustrated in FIG. 39; and
[0428] FIG. 42 illustrates the further results from exemplary primary T-cell run timeline illustrated in FIG. 40.
DETAILED DESCRIPTION
[0429] The described example embodiment relates to an apparatus, a system and a method. They primarily relate to processes in cell and/or gene therapy but are not limited thereto.
[0430] Certain terminology is used in the following description for convenience only and is not limiting. The words upper, lower, upwardly and downwardly designate directions in the drawings to which reference is made and are with respect to the described component when assembled and mounted. The words inner, inwardly and outer, outwardly refer to directions toward and away from, respectively, a designated centerline or a geometric center of an element being described (e.g., a central axis), the particular meaning being readily apparent from the context of the description. Further, the terms proximal (i.e., nearer to) and distal (i.e., away from) designate positions relative to a body or a point of attachment.
[0431] Further, as used herein, the terms connected, affixed and the like are intended to include direct connections between two members without any other members interposed therebetween, as well as, indirect connections between members in which one or more other members are interposed therebetween. The terminology includes the words specifically mentioned above, derivatives thereof, and words of similar import.
[0432] Further, unless otherwise specified, the use of ordinal adjectives, such as, first, second, third etc., merely indicate that different instances of like objects are being referred to and are not intended to imply that the objects so described must be in a given sequence, either temporally, spatially, in ranking or in any other manner. Like reference numerals are used to depict like features throughout.
[0433] FIGS. 3(a) to 3(c) illustrate a first embodiment of an actuation mechanism 100 according to the disclosure. The actuation mechanism 100 includes a moveable plate 102 and a base plate 106. The moveable plate 102 is axially translatable by virtue of a plurality of linear actuators 104. The base plate 106 may also be rotatable about an axis by one or more actuators (not shown).
[0434] FIG. 4 illustrates a second embodiment of an actuation mechanism 200. The actuation mechanism 200 includes a moveable plate 202 and a linear actuator 204. The linear actuator 204 acts upon a lower side, or surface, of the moveable plate 202 so as to raise and lower the moveable plate 202, in use.
[0435] FIGS. 5(a) and 5(b) illustrate another embodiment of an actuation mechanism 300. The actuation mechanism 300 includes a moveable plate 302 and a base plate 306. The base plate 306 is connected to the moveable plate 302 by a linkage 304. The linkage 304 may be a series of pivotally connected rods, as shown in FIGS. 5(a) and 5(b). The actuation mechanism 300 may include one or more actuators (not shown) for raising or lowering the moveable plate 302, with respect to the base plate 306, in use. Furthermore, the actuation mechanism 300 may include one or more actuators (not shown) for rotating the base plate 306 about an axis, as indicated by the arced arrow in FIGS. 5(a) and 5(b). It is noted that there may be one or more actuators, which may both raise and lower the moveable plate 302, and rotate the base plate 306 about an axis.
[0436] FIG. 6 illustrates another embodiment of an actuation mechanism 400. The actuation mechanism 400 includes a moveable plate 402 and a base plate 406. The moveable plate 402 is connected to the base plate 406 by a pivotable rod 408. There is also provided a linear actuator 404, which acts upon the lower side, or surface, of the moveable plate 402. There may be any number of linear actuators 404. In the example where a number of linear actuators 404 are presented, when the linear actuators 404 act cooperatively, i.e., simultaneously, the moveable plate 402 is axially translated, that is raised or lowered. When the linear actuators 404 do not act cooperatively, i.e., not simultaneously, or out of timing, the moveable plate 402 is rocked along one or more axes, depending upon the number of linear actuators 404.
[0437] FIG. 7 illustrates another embodiment of an actuation mechanism 500. The actuation mechanism 500 includes a moveable plate 502 and a base plate 506. The moveable plate 502 and the base plate 506 are connected by a central rod 514 extending axially from the base plate 506 toward a pivot point 512 of the moveable plate 502. There is also provided a series of springs 504a, 504b. Any number of springs 504a, 504b may be used. The springs 504a, 504b bias the moveable plate 502 toward the base plate 506. The springs 504a, 504b may be tension springs. Furthermore, there is provided a connecting rod 508, extending from the central rod 514 and terminating in a wheel 510. The central rod 514 may be driven by an actuator or a motor such that the wheel 510 is caused to rotate about the central rod 514. In this way, the moveable plate 502 is caused to rotate about two axes perpendicular to one another so as to provide a swirling motion of a container on the moveable plate 502.
[0438] FIG. 8 illustrates another embodiment of an actuation mechanism 600. The actuation mechanism 600 includes a moveable plate 602 and a base plate 606. The moveable plate 602 is connected to the base plate 606 by a pivotable rod 610. The base plate 606 is provided with electromagnets 608a, 608b on the outer edge. The moveable plate 602 is provided with permanent magnets 604a, 604b on the outer edge. The electromagnets 608a on the base plate 606 align with the permanent magnets 604a on the moveable plate 602. The electromagnets 608b on the base plate 606 align with the permanent magnets 604b on the moveable plate 602. In use, the electromagnets 608a, 608b are turned on and off sequentially such that the electromagnets 608a, 608b sequentially attract and repel the permanent magnets 604a, 604b on the moveable plate 602. The interaction between the electromagnets 608a, 608b on the base plate 606 and the permanent magnets 604a, 604b on the moveable plate 602 cause the moveable plate 602 to pivot about the pivotable rod 610. In some examples, the magnetic forces acting on the moveable plate 602 cause the moveable plate 602 to move about the base plate 606 in a wave motion.
[0439] FIG. 9 illustrates another embodiment of an actuation mechanism 700. The actuation mechanism 700 includes a moveable plate 702 and a base plate 706. The base plate 706 is connected to the moveable plate 702 by a linkage 704. The linkage 704 is a series of pivotally connected rods, arranged in a scissor lift configuration. The actuation mechanism 700 may be provided with one or more actuators (not shown) that raise and lower the moveable plate 702, with respect to the base plate 706, in use. That is, the distance between the base plate 706 and the moveable plate 702 may be changed by raising and lowering the moveable plate 702, with respect to the base plate 706 by use of the one or more actuators.
[0440] FIGS. 10(a) to 10(e) illustrate another embodiment of an actuation mechanism 800. The actuation mechanism 800 includes a moveable plate 802 and a base plate 806. The base plate 806 is connected to the moveable plate 802 by a central rod 810 extending axially from the base plate 806 toward a pivot point 812 of the moveable plate 802. A first cam member 804a is provided on the base plate 806 and in contact with the moveable plate 802. A second cam member 804b is provided on an opposite side of the central rod 810, distal the first cam member 804a. The first cam member 804a is driven by one or more motors (not shown). The second cam member 804b is driven by one or more motors (not shown). The first cam member 804a is driven by the one or more motors to rotate about its pivotal axis. The second cam member 804b is driven by the one or more motors to rotate about its pivotal axis. In use, the first cam member 804a is driven such that a surface thereof engages the moveable plate 802 such that the moveable plate 802 moves about the pivot point 812. The second cam member 804b may be driven either instead of or in addition to the first cam member 804a such that a surface thereof engages the moveable plate 802 such that the moveable plate 802 moves about the pivot point 812. The first cam member 804a may be driven independently of the second cam member 804b.
[0441] In some examples, the first cam member 804a and the second cam member 804b are driven simultaneously. The cam members 804a, 804b may have a non-circular profile such that the moveable plate 802 moves non-linearly. As shown in this particular example, the first cam member 804a is arranged to rotate out of phase in respect of the second cam member 804b in order to raise one side of the moveable plate 802 and lower the other side of the moveable plate 802. As will be described, the cam members 804a, 804b may be provided with an angled surface or a non-circular profile, to tilt the moveable plate 802 from one side to the other during use. Examples of the cam members 804a, 804b are illustrated in FIGS. 10(d) and 10(e). FIG. 10(d) shows a cam member 804a having a circular profile. FIG. 10(e) shows a cam member 804a having an elongated circular profile. As shown in FIG. 10(b), the cam members 804a, 804b may additionally be provided with tension springs 808a, 808b that provides a resilient force between the base plate 806 and the moveable plate 802. FIG. 10(c) illustrates an example in use, where the first cam member 804a and the 804b are mirrored with respect to each other. The upper surface of the cam members 804, 804b are level such that the moveable plate 802 positioned horizontally.
[0442] FIG. 11 illustrates another embodiment of an actuation mechanism 900. The actuation mechanism 900 includes a moveable plate 902 and a base plate 906. The moveable plate 902 and the base plate 906 are connected by a central hub 914 extending axially from the base plate 906 toward a pivot point 912 of the moveable plate 902. A series of springs 904a, 904b are provided. Any number of springs 904a, 904b may be used. The springs 904a, 904b bias the moveable plate 902 toward the base plate 906. In this example, the springs 904a, 904b are tension springs. A connecting rod 908 is provided, extending radially from the central hub 914 and terminating in a wheel 910. The central hub 914 may be driven by an actuator or a motor such that the wheel 910 is caused to rotated about the central hub 914. In this way, the moveable plate 902 is caused to rotate about two axes perpendicular to one another so as to provide a swirling motion of a container on the moveable plate 902. In some examples, the wheel 910 is driven by a motor and moves around in a circular motion. This moves the moveable plate 902 in a symmetrical wave motion.
[0443] FIG. 12 illustrates another embodiment of an actuation mechanism 1000. The actuation mechanism 1000 includes a moveable plate 1002 and a base plate 1006. The moveable plate 102 and the base plate 1006 are connected by a central hub 1014 that extends axially from the base plate 1006 toward a pivot point 1012 of the moveable plate 1002. Springs 1004a, 1004b are provided to bias the moveable plate 1002 toward the base plate 1006. In this embodiment, a central hub 1014 is provided on the base plate 1006. The central hub 1014 is coupled to a support plate 1010. A linear actuator 1008 is provided between the support plate 1010 and the base plate 1006. The linear actuator 1008 acts upon a lower side, or surface, of the moveable plate 1002 so as to raise and lower the moveable plate 1002, in use. The support plate 1010 may be rotationally driven by a motor (not shown).
[0444] FIGS. 13(a) and 13(b) illustrate another embodiment of an actuation mechanism 1100. The actuation mechanism 1100 includes a moveable plate 1102 and a base plate 1106. A motor 1114 is positioned on the base plate 1106 and is operably coupled to a support plate 1104. The support plate 1104 is positioned between the base plate 1106 and the moveable plate 1102. The motor 1114 operates to rotate the support plate 1104, with respect to the base plate 1106. In some examples, the motor 1114 is raised and lowered with respect to the base plate 1106. The support plate 1104 is connected to the moveable plate 1102 by a linkage 1108. The linkage 1108 may be a series of pivotally connected rods. The linkage 1108 may be enclosed by a housing, such as a bellows-based housing, so as to prevent access to the linkage 1108 during use. The actuation mechanism 1100 includes an actuator 1112 for raising or lowering the moveable plate 1102, with respect to the support plate 1104, in use. In this particular embodiment, a cam system is provided. The cam system has a first cam member 1110a and a second cam member 1110b. The cam members 1110a, 1110b are integrated with the linkages 1108 in this example. However, the cam members 1110a, 1110b may in addition or instead be integrated with the actuator 1112. The cam members 1110a, 1110b are drive such that a surface thereof engages the moveable plate 1102. This moves the moveable plate 1102 with respect to the base plate 1106.
[0445] FIGS. 14(a) and 14(b) illustrate another embodiment of an actuation mechanism 1200. The actuation mechanism 1200 includes a moveable plate 1202 and a base plate 1206. A motor 1216 is positioned on the base plate 1206 and is operably coupled to a support plate 1204. The support plate 1204 is positioned between the base plate 1206 and the moveable plate 1202. The motor 1216 operates to rotate the support plate 1204, with respect to the base plate 1206. In some examples, the motor 1216 is raised and lowered with respect to the base plate 1206. The support plate 1204 is connected to the moveable plate 1202 by a linkage 1214. The linkage 1214 may be enclosed by a housing, such as a bellows-based housing, so as to prevent access to the linkage 1214 during use. The linkage 1214 may be a series of pivotally connected rods. In use, the linkage 1214 moves the moveable plate 1202 with respect to the support plate 1204, and thus the base plate 1206. In this particular embodiment, the actuation mechanism 1200 includes a central hub 1210 extending axially toward the moveable plate 1202. Moreover, there is provided a connecting rod 1212, extending radially from the central hub 1210 and terminating in a linear actuator 1208. In use, the central hub 1210 is driven by an actuator or a motor (not shown) such that the linear actuator 1208 is caused to be driven to provide a swirling motion, once the linear actuator 1208 is actuated, for a container placed on top of the moveable plate 1202.
[0446] FIG. 15 illustrates another embodiment of an actuation mechanism 1300. The actuation mechanism 1300 includes a moveable plate 1302 and three linkages: a first linkage 1304a, a second linkage 1304b and a third linkage 1304c. The linkages 1304a, 1304b, 1304c extend from the bottom surface of the moveable plate 1302. The first linkage 1304a extends from the moveable plate 1302 at a first pivot point 1310a toward a base plate 1306a. The second linkage 1304b extends from the moveable plate 1302 at a second pivot point 1310b toward a base plate (not shown). The third linkage 1304c extends from the moveable plate 1302 at a third pivot point 1310c toward a base plate 1306b. A first actuator 1308a is positioned between the first linkage 1304a and the base plate 1306a. A second actuator 1308b is positioned between the second linkage 1304b and the base plate (not shown). A third actuator 1308c is positioned between the third linkage 1304c and the base plate 1306b. In use, the actuators 1308a, 1308b, 1308c are driven sinusoidally out of phase from each other to produce a wave effect on the moveable plate 1302. In some examples, the actuators 1308a, 1308b, 1308c are linear actuators that slide along rails such that the pivot points move in synchronization to move the moveable plate 1302 in a wave motion.
[0447] FIG. 16(a) illustrates another embodiment of an actuation mechanism 1400. The actuation mechanism 1400 has a moveable plate 1402 and a base plate 1406. In this particular embodiment, three motors 1408a, 1408b, 1408c are provided. Three linkages: a first linkage 1404a, a second linkage 1404b and a third linkage 1404c, are also provided extending from the moveable plate 1402. The motors 1408a, 1408b, 1408c are each used to drive a respective linkage 1404a, 1404b, 1404c. More specifically, a first motor 1408a drives a first linkage 1404a. A second motor 1408b drives a second linkage 1404b. A third motor 1408c drives a third linkage 1404c. The linkages 1404a, 1404b, 1404c are driven in coordination to move the moveable plate 1402 in a wave motion. Referring to FIGS. 16(b) and 16(c), a holding element 1450 is provided. A container 1480 having a base section is provided. The container 1480 will be described in more detail with reference to the later drawings.
[0448] In particular, it is noted that the apparatus includes the actuation mechanism 1400, the moveable plate 1402 and the holding element 1450. A system further includes the container 1480.
[0449] More specifically, the base of the container 1480 is placed on top of the moveable plate 1402 such that the container 1480 is arranged between the holding element 1450 and the moveable plate 1402. The container 1480 has a compressible wall. When the moveable plate 1402 is moved using the aforementioned actuation mechanism 1400, the top of the container 1480 is caused to be pushed against the holding element 1450. This compresses the wall of the container 1480.
[0450] FIGS. 17(a) and 17(b) illustrate another embodiment of an actuation mechanism 1500. The actuation mechanism has a moveable plate 1502 and two linear actuators 1504a, 1504b. A first linear actuator 1504a is provided on a first side of the moveable plate 1502. A second linear actuator 1504b is provided on an opposite site of the moveable plate 1502. The linear actuators 1504a, 1504b are positioned to act upon the bottom surface of the moveable plate 1502. There may be any number of linear actuators 1504. When the linear actuators 1504 act cooperatively, i.e., simultaneously, the moveable plate 1502 is axially translated. When the linear actuators 1504 do not act cooperatively, i.e., not simultaneously, or out of timing, the moveable plate 1502 is rocked along one or more axes, depending upon the number of linear actuators 1504. In this particular embodiment, a further linear actuator 1504c, which is a larger linear actuator 1504c in this example, is provided at the bottom of the actuation mechanism, below the linear actuators 1504a, 1504b. In use, the larger linear actuator 1504c is provided to be raised from an upper position, shown in FIG. 17(a), to a lower position, shown in FIG. 17(b).
[0451] FIGS. 18(a) to 18(d) illustrate another apparatus 1600 according to the disclosure. The apparatus 1600 includes a moveable plate 1602 and a base plate 1606. The moveable plate 1602 and the base plate 1606 are operably coupled by a first linkage 1604a and a second linkage 1604b. The first linkage 1604a is driven by a first motor 1608a, connected to a first gearbox 1610a and to the first linkage 1604a. The second linkage 1604b is driven by a second motor 1608b, connected to a second gearbox 1610b and to the second linkage 1604b. Each motor 1608a, 1608b, specifically the gearboxes 1610a, 1610b, is coupled to a motor pivot shaft 1612a, 1612b, which is in turn coupled to a motor coupling 1614a, 1614b for driving the linkages 1604a, 1604b.
[0452] As best shown in FIG. 18(b), the first linkage 1604a is upstanding from the base plate 1606 and coupled to the moveable plate 1602 at one end. The second linkage 1604b is upstanding from the base plate 1606 and coupled o the moveable plate 1602 at another, opposing, end. Specifically, the first linkage 1604a includes a pivot bar 1616a connected to an underside surface of the moveable plate 1602. The second linkage 1604b also includes a pivot bar 1616b connected to an underside surface of the moveable plate 1602. The moveable plate 1602 is also connected to a central pivot clamp 1618, which is pivotable about a central pivot bar 1620. Furthermore, each linkage 1604a, 1604b includes a hard stop 1622a, 1622b, defining the lowest point, or the lowest plane, in which the moveable plate 1602 can reach.
[0453] Referring now to, in particular, FIG. 18(c), the apparatus 1600 further includes a mount for a sensor 1624, which can receive a sensor 1626 therein, the sensor 1626 including a sensor cable 1628. The sensor cable 1628 communicatively connects the sensor 1626 to a controller (not shown). The controller (not shown) may further be communicatively connected to the motors 1608a, 1608b.
[0454] Further, the apparatus 1600 includes a telescopic rail 1630 mounted upon a rail mounting plate 1631, the telescopic rail 1630 moveable in a longitudinal direction and operably connected to the moveable plate 1602. Thus, the moveable plate 1602 is moveable along the telescopic rail 1630.
[0455] Furthermore, each linkage 1604a, 1604b includes a hinge 1632 and bearings 1634. There is generally provided four pivot bearing blocks 1636, 1638, 1640 (fourth not shown) in which the linkages 1604a, 1604b extend through and are supported thereon.
[0456] In use, the apparatus 1600 is arranged to rotate the moveable plate 1602 about the central pivot bar 1620, thereby imparting a wave motion, or a rocking motion, to a container engaged by the moveable plate 1602. Additionally, in use, the moveable plate 1602 can be translated along a longitudinal axis, centrally and perpendicularly disposed through the moveable plate 1602, thereby imparting a compression motion to a container engaged by the moveable plate 1602.
[0457] The user may operate the apparatus 1600 according to a pre-set program stored within a controller (not shown), which, when it executes a series of operating instructions, performs the desired motion. Alternatively, the user may input parameters into the controller (not shown) to operate the apparatus 1600 according to a desired range of motion.
[0458] Although two linkages 1604a, 1604b are shown in the present embodiment, any number of linkages may be used. Additionally, although the linkages 1604a, 1604b are driven by motors 1608a, 1608b, there may alternatively by driven by other mechanisms such as actuators, for example, linear actuators.
[0459] FIG. 19 illustrates an example of a linear actuator 1700. The linear actuator 1700 may act upon, that is engage, a portion or a surface of any moveable plate as described herein. Alternatively, or additional in embodiment in which a plurality of linear actuators are provided, the linear actuator 1700 may be coupled to a portion or a surface of any moveable plate as described herein.
[0460] FIGS. 20(a) and 20(b) illustrate a container before and after a swirling motion, respectively. In particular, FIG. 20(a) shows a container 1800 including a central origin 1802 in which a longitudinal axis, extending perpendicularly to a base section of the container 1800, transects the plane of the base section of the container. Such a longitudinal axis is further described in relation to FIGS. 28(a) to 30(c). The container 1800 includes water and several droplets of food coloring 1804 to demonstrate the various agitation methods described herein. As can be seen in FIG. 20(b), after imparting a swirling motion to the water and the food coloring 1804 by pivoting the container 1800 about the central origin 1802, the food coloring begins to mix, as indicated by 1806. Continual swirling provides full dispersion of the food coloring 1804 within the water in the container 1800. This provides an illustration of the mixing of a cell processing medium within the container 1800, in use.
[0461] FIGS. 21(a) and 21(b) illustrate a container before and after a wave motion, respectively. In particular, FIG. 21(a) shows a container 1900 including an axis 1902 running perpendicular to a longitudinal axis as described in relation to FIGS. 20(a) and 20(b), and running within the plane of the base of the container 1900. The container 1900, like the container 1800 of FIGS. 20(a) and 20(b), includes water and a food coloring to demonstrate the various agitation methods described herein. As can be seen in FIG. 21(b), after imparting a wave motion to the water and the food coloring by rotating, tilting, or pivoting, the container 1900 about the axis 1902 (FIG. 21(a)), the food coloring begins to mix, particularly toward an wall element of the container 1900. In particular, there may be constructive interference, as indicated by 1904, toward the wall element of the container 1900, thereby aiding mixing of the water and the food coloring. Continual wave agitation provides full dispersion of the food coloring within the water in the container 1900. This provides an illustration of the mixing of a cell processing medium within the container 1900, in use.
[0462] FIGS. 22(a) to 22(c) illustrates a container having water and a food coloring therein for simulating the mixing of a cell processing medium within the container. FIG. 22(a) illustrates the container in which food coloring has been dropped into the water at time t=0 seconds. FIG. 22(b) illustrates the container in which the food coloring begins to mix with the water at time after t=0. FIG. 22(c) illustrates the container in which the food coloring is mixed with the water at time after t=0 and after that shown in FIG. 22(b). Throughout FIG. 22, a swirling motion, as described in relation to FIGS. 20(a) and 20(b), is imparted to the water and the food coloring within the container, at 20 revolutions per minute having a water content of 500 mL.
[0463] FIGS. 23(a) to 23(c) illustrates a container having water and a food coloring therein for simulating the mixing of a cell processing medium within the container. FIG. 23(a) illustrates the container in which food coloring has been dropped into the water at time t=0 seconds. FIG. 23(b) illustrates the container in which the food coloring begins to mix with the water at a time after t=0 seconds. FIG. 23(c) illustrates the container in which the food coloring is mixed with the water at a time after t=0 seconds and after that shown in FIG. 23(b). Throughout FIG. 23, a swirling motion, as described in relation to FIGS. 20(a) and 20(b), is imparted to the water and the food coloring within the container, at 30 revolutions per minute having a water content of 500 mL.
[0464] FIGS. 24(a) to 24(c) illustrates a container having water and a food coloring therein for simulating the mixing of a cell processing medium within the container. FIG. 24(a) illustrates the container in which food coloring has been dropped into the water at time t=0 seconds. FIG. 24(b) illustrates the container in which the food coloring begins to mix with the water at time after time t=0 seconds. FIG. 24(c) illustrates the container in which the food coloring is mixed with the water at time after t=0 seconds and after that shown in FIG. 24(b). Throughout FIG. 24, a swirling motion, as described in relation to FIGS. 20(a) and 20(b), is imparted to the water and the food coloring within the container, at 60 revolutions per minute having a water content of 500 mL.
[0465] Thus, as can be seen by comparing FIGS. 22(c), 23(c) and 24(c), at relatively high volumes of fluid, such as 500 mL as demonstrated, a swirl motion does not appear to adequately mix the food coloring within the water. This is a good model of cell processing media within a container.
[0466] As shown in FIGS. 25(a) and 25(b) a container having water and a food coloring therein for simulating the mixing of a cell processing medium within the container. FIG. 25(a) illustrates the container in which food coloring has been dropped into the water at time t=0 seconds. FIG. 25(b) illustrates the container in which the food coloring begins to mix with the water at time t=60 seconds. Unlike FIGS. 22(a) to 24(c), through FIGS. 25(a) and 25(b) a wave motion, as described in relation to FIGS. 21(a) and 21(b), is imparted to the water and the food coloring within the container, at 20 revolutions per minute having a water content of 50 mL.
[0467] Thus, as can be seen by comparing FIG. 25(b) with FIGS. 22(c), 23(c) and 24(c), a swirling motion is more suitable for mixing the contents of the container at lower fluid volumes, even at low revolutions per minute. Therefore, a swirling motion may be preferably for small volumes of fluid in cell processing.
[0468] FIGS. 26(a) to 26(c) illustrates a container having water and a food coloring therein for simulating the mixing of a cell processing medium within the container. FIG. 26(a) illustrates the container in which food coloring has been dropped into the water at time t=0 seconds. FIG. 26(b) illustrates the container in which the food coloring begins to mix with the water at a time after t=0 seconds. FIG. 26(c) illustrates the container in which the food coloring is mixed with the water at a time after t=0 and after that shown in FIG. 26(b). Throughout FIG. 26, a linear compression motion, that is a compression along a longitudinal axis as described in further detail below, is imparted to the water and the food coloring within the container, at 20 cycles per minute having a water content of 500 mL. As can be seen in FIG. 26(c), at low cycles per minute, mixing appears to be largely due to diffusion of the food coloring in the water.
[0469] FIGS. 27(a) to 27(c) illustrates a container having water and a food coloring therein for simulating the mixing of a cell processing medium within the container. FIG. 27(a) illustrates the container in which food coloring has been dropped into the water at time t=0 seconds. FIG. 27(b) illustrates the container in which the food coloring begins to mix with the water at a time after t=0 seconds. FIG. 27(c) illustrates the container in which the food coloring is mixed with the water at a time after t=0 seconds and after that shown in FIG. 27(b). Throughout FIG. 27, a linear compression motion, that is a compression along a longitudinal axis as described in further detail below, is imparted to the water and the food coloring within the container, at 40 cycles per minute having a water content of 500 mL. As can be seen in FIG. 27(c), at increased cycles per minute, mixing is improved.
[0470] FIGS. 28(a) to 28(c) illustrates a container having water and a food coloring therein for simulating the mixing of a cell processing medium within the container. FIG. 28(a) illustrates the container in which food coloring has been dropped into the water at time t=0 seconds. FIG. 28(b) illustrates the container in which the food coloring begins to mix with the water at a time after t=0 seconds. FIG. 28(c) illustrates the container in which the food coloring is mixed with the water at time after t=0 seconds and after that shown in FIG. 28(b). Throughout FIG. 28, a linear compression motion, that is a compression along a longitudinal axis as described in further detail below, is imparted to the water and the food coloring within the container, at 80 cycles per minute having a water content of 500 mL. As shown in FIG. 28(b), occasionally, large bubbles are created during the linear compression, which can aid in aerating the mixture, which can be useful in cell processing methods. As shown in FIG. 28(c), good mixing is achieved for higher volumes of fluid at larger cycles per minute.
[0471] Thus, it has been found that at small volumes, particularly less than 100 mL, a swirling motion is suitable for adequately mixing a fluid in the container. Moreover, it has been found that at larger volumes, particularly 100 mL or greater, a linear compression motion is suitable for adequately mixing a fluid in the container. Thus, an agitation mechanism should be able to provide both ranges of motion, so that any volume of fluid may be adequately mixed.
[0472] FIGS. 29(a) and 29(b) illustrate a container according to the disclosure. The container 2000 includes a top section 2002, a base section 2004 extending in parallel to the top section 2002, and a wall element 2006 therebetween. The wall element 2006 is compressible along the longitudinal axis L of the container 2000. The wall element 2006 may comprise one or more Z-folds 2008, or may form a concertina, such that it is compressible along the longitudinal axis L. FIG. 29(a) illustrates the container 2000 prior to compression. FIG. 29(b) illustrates the container 2000 after compression of a portion of the wall element 2006. Generally, each of the top section 2002 and the base section 2004 generally define a plane, each plane being parallel to one another and substantially perpendicular to the longitudinal axis L.
[0473] FIGS. 30(a) to 30(e) illustrate an apparatus 2100 including a container 2000, according to the disclosure. The apparatus 2100 includes a holding element 2110 and a moveable plate 2120. The moveable plate 2120 can be any moveable plate as described herein, and moveable by any actuation mechanism as described herein. The holding element 2110 holds a top section of the container 2000 within a first plane P1, and the moveable plate 2120 forms a second plane P2, substantially parallel to the first plane P1, for moving the base section of the container 2000. Prior to operation, i.e., actuation and moving of the moveable plate 2120, the first and second planes P1, P2 extend substantially horizontally, and substantially perpendicular to the longitudinal axis L.
[0474] As shown in FIG. 30(a), the container 2000 may be compressed along the longitudinal axis L, such that the first plane P1 remains stationary and such that the moveable plate 2120, and thus the second plane P2, is caused to move. Compression along the longitudinal axis L may cause mixing of media within the container 2000 as described above. In the example show, a 50 ml transduction may take place. The liquid may have a depth of approximately 2.75 mm.
[0475] As shown in FIG. 30(b), the container 2000, containing media 2102, may be expanded, or decompressed, along the longitudinal axis L, such that the first plane P1 remains stationary and such that the moveable plate 2120, and thus the second plane P2, is caused to move. Expansion along the longitudinal axis L may be beneficial in the cell processing method, for example, for cell cultivation or growth. In the example shown, a 500 mL expansion may take place. The liquid may have a depth of approximately 27 mm.
[0476] As shown in FIG. 30(c), the container 2000 may be subjected to further compression, thus agitating the media 2102 therein, along the axis L as indicated by the arrow. This causes the container 2000, specifically the wall element thereof, to be compressed along the longitudinal axis L. That is, the first plane P1 remains stationary and the moveable plate 2120, and the second plane P2, are moved.
[0477] As shown in FIG. 30(d), as the headspace within the container 2000 above the media 2102 is reduced, media 2102 residing within the wall element of the container 2000 is forced inwardly (directional arrows 2130 and 2120), toward the longitudinal axis L. This motion causes agitation within the container 2000 of the media 2102. This may be beneficial to a number of steps in the cell processing method.
[0478] As shown in FIG. 30(e), as the container 2000 is expanded, or decompressed, along the longitudinal axis L, media 2104, 2106, which has collected within the wall element of the container 2000 during compression, is caused to flow, drop or drip, into the bulk of the media 2102, thereby causing further agitation within the container. This may be beneficial to a number of steps in the cell processing method.
[0479] The compression may force medium to be compression out of the folds as the container is raised and lowered repeatedly. Compression pauses to a set period of time may also be providedthat is, a compression may be held, or stopped, such that the container is in a compressed state for a predetermined period of time. This allows cell sedimentation before medium exchange. The compression steps may result in excellent mixing and gaseous transfer. Such a process may also be scalable. Thus, there may be a number of advantages associated with such a compression process described herein for cell processing methods.
[0480] FIGS. 31(a) to 31(c) illustrate a further apparatus 2200 of the disclosure. The apparatus 2200 includes a holding element 2210 for receiving and holding a top section of the container 2000 within a first plane P1. The apparatus 2200 further includes a moveable plate 2220 for acting upon, or engaging, a base section of the container 2000, the moveable plate 2220 defining a second plane P2. The first plane P1 and the second plane P2 are substantially parallel and extend substantially perpendicularly to the longitudinal axis L.
[0481] As shown in FIG. 31(a), a static phase or step of a method is shown, in which the container 2000 is not agitated in any way. This may be beneficial to a number of steps in the cell processing method.
[0482] As shown in FIG. 31(b), an agitation phase or step of a method is shown, in which the container 2000 is agitated so as to cause agitation, or turbulence, of media within the container 2000. In this particular example, the agitation phase includes rotating the moveable plate 2220 about at least one axis within the plane P2, for example, the axis A indicated in FIG. 31(a). The plane P2 moves as the moveable plate 2220 moves. Thus, as the moveable plate 2220 moves, the plane P2, or a portion thereof, moves toward the plane P1, which remains stationary. Thus, a distance between at least a portion of the plane P2 and the plane P1 is decreased. The movement of the moveable plate 2220 about the axis A causes a portion of the wall element of the container 2000 to be compressed, as shown in FIG. 31(b). As the moveable plate 2220 rotates, or tilts, in the opposition direction, an opposing portion of the wall element of the container 2000 is compressed. Thus, a wave motion, as described above, may be effected upon the media within the container 2000. Furthermore, the moveable plate 2220 may be pivotable about an origin, in which the longitudinal axis L intersects the second plane P2, such that the origin remains stationary and all other points move. In this example, a swirling motion, as described above, may be effected upon the media within the container 2000. In other examples, the moveable plate 2220 may be rotatable or arranged to be tilted about any number of axes within the second plane P2 to cause any desired agitative effect on the media within the container 2000.
[0483] As shown in FIG. 31(c), a compression phase or step of a method is shown, in which the container 2000, specifically the wall element, is compressed along the longitudinal axis L such that the first plane P2 is caused to move closer toward the first plane P1. Thus, as shown in FIG. 31(c), the headspace above the media within the container 2000 may be changed, for example, air may be pushed out of the container 2000. Further compression may cause agitation as described above.
[0484] FIG. 32 illustrates a mold or an insert 2300 for producing a container as described herein. The mold 2300 may be suitable for blow-molding. As such, the container may be formed by blow-molding. The mold 2300 includes a series of protrusions, or ridges, 2302, arranged to provide the series of Z-folds, or the concertina arrangement, of the containers described herein.
[0485] FIGS. 33(a) to 33(d) illustrate an example of a container 2400, a platform 2410 and other components that may be used in combination with the apparatuses or systems described herein. FIG. 33(d) shows a container 2400 having a top section that is substantially open thereby forming a fluid passageway, and a base section that is substantially closed. There is a wall element extending between the top section and the base section as described in relation to FIGS. 29(a) and 29(b). The wall element may comprise a compressible portion, for example, Z-folds as shown in FIGS. 33(b) and 33(d). The container 2400 may form a bellows, for example, a 4-fold bellows, or a concertina arrangement.
[0486] Referring further to FIG. 33(d), the container 2400 is attachable to a platform 2410, which may be regarded as a cell processing platform, at an underside thereof. There may also be provided a clip 2412, which may form the holding element as described herein. The clip 2412 may be provided in two halves as shown. The container 2400 may be sealed by way of an O-ring 2414 that is urged between the top section of the container 2400 and the underside of the platform 2410. The clip 2412 may be coupled to the platform 2410 by way of bolts 2416 and corresponding nuts 2418.
[0487] The platform 2410 may have any number of features. For example, the platform 2410 may have a series of fluid inlets having attachment elements for attaching various pieces of equipment. The platform 2410 may have one or more, preferably one, fluid outlet in fluid communication with the fluid inlets. For example, as shown in FIG. 33(d), the attachment elements of the inlets of the platform 2410 may be coupled to a secondary container 2420 having a neck 2422 including a screw thread for screwing to a corresponding screw thread of an attachment mechanism 2424 of the platform 2410. The attachment mechanism 2424 may also include an O-ring 2426 to ensure a fluid or liquid tight seal. The platform 2410 may also include a male connector element 2428 for connecting the platform 2410 to one or more tubes or other pieces of equipment. The male connector element 2428 is fluid coupled to a fluid inlet of the platform 2410 by tubing 2430, for example, flexible tubing, held in place by a cable tie 2432. A filter 2434 may also be present between the flexible tubing 2430 and the male connector element 2428. The platform 2410 may also include tubing 2436 coupled to an inlet of the platform 2410 for connecting to other pieces of equipment. The tubing 2436 may be flexible tubing. The tubing 2436 is coupled to the fluid inlet of the platform 2410 by a cable tie 2438. The platform 2410 may also include a cap 2440 for covering one or more elements, for example, a sampling element 2442 or the inlets, of the platform 2410.
[0488] In use, the secondary container 2420 acts as a breathing container for the container 2400. That is, as the container 2400 is compressed or decompressed or otherwise moved by, for example, a moveable plate of the disclosure, the secondary container 2420 allows for the compensation of pressure increases and/or decreases in the container 2400. In particular, as the container 2400 is acted upon, at its base section by the moveable plate, fluid, for example, air, may be forced out of the container 2400, through the platform 2410, and into the secondary container 2420. Likewise, if the container 2400 is pulled downwardly, or otherwise decompressed, then fluid, for example, air, may be drawn into the container 2400, through the platform 2410 from the secondary container 2420. Thus, the secondary container 2420 acts as a breathing container in that it accounts for pressure changes in the container 2400.
[0489] FIGS. 34(a) to 34(d) illustrate another apparatus 2500, and components thereof, in accordance with the disclosure. As best shown in FIGS. 34(a) and 34(b), the apparatus 2500 includes a moveable plate 2502, formed with two protruding arms 2502a, 2502b for frictionally engaging a side wall adjacent a base of a container 2580. The apparatus 2500 is depicted without the holding element, for clarity only, though this is discussed in further detail in relation to FIGS. 35(a) to 35(e) below.
[0490] The apparatus includes a linkage 2504 connecting the moveable plate 2502 to a first motor 2508a, a second motor 2508b and a third motor 2508c. Each motor has respective gearboxes 2510a, 2510b, 2510c. The first motor 2508a is arranged to rotate the moveable plate 2502 about an axis formed within a plane thereof. The second and third motors 2508b, 2508c, are arranged to move the moveable plate 2502 in a longitudinal direction as described in more detail below.
[0491] Specifically, the linkage 2504 illustrated includes a first longitudinal rail 2504a and a second longitudinal rail 2504b. Each longitudinal rail 2504a, 2504b is generally formed as a pair of complementary rails in this particular embodiment. The linkage 2504 further includes a crank housing 2512, including a crank 2514 driven by a shaft 2516, which is connected to the second motor 2508b, which is slidable along the first longitudinal rail 2504a in an upward and/or downward direction.
[0492] The linkage 2504 further includes a slider portion 2520, connected to the moveable plate 2502, which is slidable along the second longitudinal rail 2504b in an upward and/or downward direction. The crank 2514 is connected to the slider portion 2520 by a connecting rod 2518. The slider portion 2520 also includes a shaft (not shown) extending therethrough so as to couple the moveable plate 2502 to the first motor 2508a.
[0493] Referring further to FIG. 34(c), the crank arrangement discussed above is illustrated in more detail. In particular, there is provided the crank 2514 driven by the shaft 2516 by the second motor (not shown). The crank 2514 is coupled to a connecting rod 2518, which terminates in the slider portion 2520. The crank 2514 has a diameter of 40 mm in the depicted example, but other diameters are equally contemplated. The crank 2514 may be replaceable, such that it can be replaced by larger or smaller diameter cranks such that the displacement of the moveable plate during use is adjusted accordingly. For example, for larger displacements, a larger diameter crank may be used. For example, for smaller displacements, a smaller diameter crank may be used.
[0494] In use, as the crank 2514 is caused to rotate, and thus the connecting rod 2518 moves, the slider portion 2520 is caused to axially move upwardly and then, as the crank 2514 continues to rotate, is caused to axially move downwardly. As such, the slider portion 2520 is moveable between a first position, or a home position, and a second position, or an elevated position, by the crank 2514. The slider portion 2520 then returns to the first position, or the home position, upon continuing rotation of the crank 2514. In this way, the crank arrangement allows for rapid upward and downward movement of the moveable plate, thus imparting a full compression and decompression cycle to a received container, during use, with each rotation of the crank 2514.
[0495] Referring further to FIG. 34(d), the third motor 2508c is illustrated in more detail. In particular, the third motor 2508c is shown as a lead screw thread motor or a ball screw thread motor, including a drivable threaded shaft 2522. The driveable threaded shaft 2522 is threadedly coupled to a corresponding screw threaded portion 2522, which forms part of the crank housing 2512 (see FIG. 34(a)). As such, as the threaded shaft 2522 is driven, the screw threaded portion 2522 of the crank housing 2512 is caused to axial move, either upwardly or downwardly depending upon the rotational direction of the driven shaft 2524. As such, the crank housing 2512 is moveable along the first longitudinal rail 2504a (see FIG. 34(a)). Thus, owing to the crank arrangement discussed above, the moveable plate 2502 (see FIG. 34(a)) is similarly moveable, along the second longitudinal rail 2504b (see FIGS. 34(b) and (c)). In this way, the lead screw or ball screw arrangement allows for slower upward and downward movement of the moveable plate.
[0496] Referring to FIGS. 34(a) to 34(d), during use, a container 2580 is received within the apparatus 2500, specifically between the moveable plate 2502 and a holding element (not shown). The first motor 2508a is caused to be actuated, so as to rotate the moveable plate 2502 about an axis within the plane of the moveable plate 2502. Furthermore, the second motor 2508b is caused to be actuated, either intermittently or continuously, so as to cause axial translation of the moveable plate 2502 along a longitudinal axis, parallel to the first and second longitudinal rails 2504a, 2504b. As discussed above, the second motor 2508b causes the shaft 2516 to rotate, thereby rotating the crank 2514. As the crank 2514 rotates, the connecting rod 2518 is caused to move, thereby axially moving the slider portion 2520 along the second longitudinal rail 2504b. Owing to the configuration of the crank arrangement, the slider portion 2520, and thus the moveable plate 2502, is caused to move axially upwardly and downwardly, thereby imparting a compression and decompression motion to the container 2580. Further, due to this configuration, the axial movement of the slider portion 2520, by the second motor 2508b, is limited to a predetermined region along the longitudinal axis. Furthermore, the third motor 2508c is caused to be actuated, so as to cause rotation of the driven shaft 2524, thereby axially translating the crank housing 2512 upwardly or downwardly, depending upon the direction of rotation of the driven shaft 2524. As such, the crank housing 2512 is caused to axially move along the first longitudinal rail 2504a, thereby imparting a slow compression or decompression motion to the container 2580.
[0497] For completeness, the person skilled in the art would readily appreciate that the motors 2508a, 2508b, 2508c can be actuated in any order, or indeed in combination with one another. The above description of the use of the apparatus 2500 is not intended to be limiting in any sense.
[0498] FIGS. 35 and 36(a) to 36(d) illustrate another apparatus 2600 of the disclosure, which operates based upon the same principle as noted in the apparatus 2500 above of FIGS. 34(a) to 34(d).
[0499] FIG. 35 illustrates the apparatus 2600 including a moveable plate 2602 operated by the linkage 2604, motors 2608a, 2608b, 2608c and gearboxes 2610a, 2610b, 2610c as described in relation to FIGS. 34(a) to 34(d). Further explanation will not be provided for brevity. The apparatus 2600 further includes a holding element 2650. The holding element 2650 is formed of an upper plate 2652, arranged to receive a top portion of a container in use, and a lower plate 2654. The moveable plate 2602 is arranged between the upper plate 2652 and the lower plate 2654. The upper plate 2652 is connected to the lower plate 2654 by a plurality of legs 2656 extending therebetween and coupled to each plate 2652, 2654 at their ends. The upper plate 2652 generally includes a recessed surface 2658 for receiving a top portion of a container in use and as described further below. The upper plate 2652 also includes a plurality of clamps 2660 arranged to clamp a top portion of a container, in use, to the upper plate 2652.
[0500] The apparatus 2600 also includes a frame 2662 formed about the actuation mechanism, including the linkage 2604, motors 2608a, 2608b, 2608c and the gearboxes 2610a, 2610b, 2610c. Although not illustrated in this drawing for clarity, the frame 2662 also includes a plurality of panels enclosing the components of the actuation mechanism, thereby preventing user access during use. Further, the panels may prevent the ingress liquid, or other cell processing components such as media, cells, or similar, into the actuation mechanism. The frame 2662 and the panels may be regarded as forming a housing for the actuation mechanism.
[0501] FIGS. 36(a) to 36(e) illustrate the apparatus 2600, described in relation to FIG. 35, having the container 2400 and platform 2410 of FIGS. 33(a) to 33(d) attached thereto. In particular, a base section of the container 2400 is engageable with the moveable plate 2602. In this particular embodiment, the base section of the container 2400 is allowed to engage, and disengage, freely with the moveable plate 2602, but in other examples the base section of the container 2400 may be fixedly coupled or attached, such as through adherence, fastening means or the like, to the moveable plate 2602. Further, the platform 2410, which is coupled to the top section of the container 2400, is received within the recessed surface 2658 (see FIG. 35) and clamped thereto by a plurality of clamps 2660. As such, the container 2400 is positioned between the moveable plate 2602 and the holding element 2650, so as to allow for compression, decompression, or other movement, of the container 2400 during use.
[0502] During use, the respective motors 2608a, 2608b, 2608c are actuated so as to cause desired movement of the container 2400, in a manner analogous to that described in relation to FIGS. 34(a) to 34(d). In particular, the first motor 2608a controls rotational movement of the base of the container 2400, about an axis formed within the plane of the moveable plate 2602, the second motor 2608b controls rapid compression/decompression of the container 2400, specifically by axially translating the base of the container 2400 toward, or away from, the holding element 2650, and the third motor 2608c controls slow compression/decompression of the container 2400, specifically by axially translating the base of the container 2400 toward or away from, the holding element 2650. The first motor 2608a thus may be useful for imparting a swirling or rocking motion to the base of the container 2400. The second motor 2608b thus may be useful for imparting a compression mixing motion, i.e., rapid compression followed by rapid decompression, to the container 2400. The third motor 2608c thus may be useful for imparting a breathing motion, for example, by causing fluid, such as air, media or the like, to be expelled from the container 2400 and into the secondary container 2420, described in further detail in relation to FIGS. 33(a) to 33(d). As such, distinct motors may be provided for the distinct, or indeed composite during use, motions imparted to the container 2400.
[0503] In some examples herein, there may be a method 2700, illustrated by FIG. 37, for cell processing. The method 2700 may include the step of providing 2710 a compressible container including a population of cells in a liquid medium. The method 2700 may include the step of statically processing 2720 the population of cells in the liquid medium within the compressible container. The method 2700 may include the step of dynamically processing 2730 the population of cells in the liquid medium within the compressible container.
[0504] The step of statically processing 2720 the population of cells in the liquid medium may comprise not subjecting the population of cells to any movement or force. That is, there may be a static phase of cell processing.
[0505] The step of dynamically processing 2730 the population of cells in the liquid medium may include agitating the population of cells in the liquid medium. The step of agitating the population of cells in the liquid medium may include imparting a wave motion to the population of cells and the liquid medium, imparting a swirling motion to the population of cells and the liquid medium, compressing the compressible container, or a combination thereof. The step of compressing the compressible container may include compressing the compressible container along a longitudinal axis that is perpendicular to a top section and a base section of the container.
[0506] There may be provided a separate step, or a step in combination with any other step, of compressing 2740 the compressible container including the cell population in the liquid medium.
[0507] Further embodiments and examples are provided below with reference to FIGS. 38 to 42.
EXAMPLES
[0508] The disclosure will now be described with reference to the following non-limiting examples, which demonstrate various embodiments of the disclosure. It is noted that the container 2400 and the platform 2410 of FIGS. 33(a) to 33(d) was utilized in combination with the apparatus 1600 of FIGS. 18(a) to 18(d) for the below examples:
Materials
[0509] The following materials were utilized: [0510] CD3+ T cells, healthy donor 1 (HD1) (Isolated from leukopak, Access Biologicals LLC) [0511] X-VIVO 15 (LZBE02-053Q, Lonza) [0512] 5% Normal human AB serum (H4522, Sigma) [0513] rhIL-2 (100 units.Math.mL.sup.?1) (202-IL-050, R&D Systems) [0514] Activation: CTS Dynabeads (3:1 bead-cell ratio) (40203D, ThermoFisher) [0515] Transduction: GFP Lentiviral vector (MOI:1) (0010VCT, Takarabio)
Methods
[0516] The following methods were utilized:
[00001] [0517] Feeding: Fed batch, volume doubled from set timepoint at end of static period (Day 3). Not dictated by cell density from sampling. Media volume additions contain fresh IL2 cytokine (100 units.Math.mL). [0518] Controls: FEP static culture bags (CellGenix VueLife) of increasing volume (see below) to replicate manual process for both static and early expansion phase with the same seeding density and feeding strategy (without agitation). [0519] Viable cell density and fold expansion: Cellometer Auto 2000, Nexcelom
Testing Run Timeline:
[0520] The general testing run timeline is shown in FIG. 38. In particular, inoculation is provided at Day 0. Lentiviral vector is added at Day 1. From Day 0 to Day 3, static and/or intermittent rocking motion is imparted to the container by the apparatus. Feeding, i.e., the supply of nutrients in media, is undertaken on Day 3, and samples also taken on Day 3 for testing. From Day 3 to Day 5, a rocking motion is imparted to the container by the apparatus. Further feeding is undertaken on Day 5, and samples also taken on Day 5 for testing. Finally, from Day 5 to Day 7, linear compression is imparted to the container by the apparatus. Further feeding is undertaken on Day 6 and samples also taken on Day 6 for testing. A final readout, such as for cell viability and the number of total viable cells, is taken at, or following, Day 7. Whilst this testing run provides a general process for culturing cells using the apparatus, further specific examples are noted below.
Example 1
[0521] For the Primary T-cell Run 1 (FIGS. 39 and 41), the cells and culture media were placed in a bioreactor, specifically a container 2400 illustrated in FIGS. 33(a) to 33(d). The cell culture process is substantially as described above in the testing run timeline. In this particular example, inoculation was provided at Day 0 at a volume of 50 mL utilizing the above materials, followed by lentiviral vector addition. A sample was taken on Day 3, and a feeding volume of 150 mL provided to the container. Further samples were taken on Days 4, 5 and 6. Further feeding, at a feeding volume of 100 mL, was provided on Day 6. Further feeding was provided on Days 7, 8, 9, and 10 at feed volumes of 100 mL, 400 mL, 100 mL and 50 mL, respectively. Samples were taken on Days 7, 8, 9, 10 and 11.
[0522] Additionally, the following process, or specifically the following agitation, steps were used: [0523] 1. Days 0 to 3 and Days 4 to 5: none (i.e., static culture) [0524] 2. Days 3 to 4: rocking motion A (60 rpm, displacement amplitude of 20 mm) [0525] 3. Days 5 to 6: rocking motion B (20 rpm, displacement amplitude of 10 mm) [0526] 4. Days 6 to 11: linear compression (60 cpm, displacement amplitude of 20 mm)
[0527] The relevant agitation steps (either 1, 2, 3 or 4) are indicated in FIGS. 39 and 41 at the appropriate time intervals. The total viable cells are illustrated in FIGS. 39 and 41 for this particular agitation regime, whilst FIG. 41 also illustrates the cell viability.
[0528] The key outcomes of the primary T-cell run 1 were that: [0529] One vessel can used for all volumes: no manual transfers were required [0530] Rocking motion A (Day 3 to 4): cell death was observed [0531] Cell yield after 4 days of compression mixing was >1?10.sup.9 [0532] Final viability was within release criteria (?80%)
Example 2
[0533] For the Primary T-cell Run 2 (FIGS. 40 and 42), the cells and culture media were placed in a bioreactor, specifically a container 2400 illustrated in FIGS. 33(a) to 33(d). The cell culture process is substantially as described above in the testing run timeline. In this particular example, inoculation was provided at Day 0 at a volume of 50 mL, followed by lentiviral vector addition. A sample was taken on Day 3, and a feeding volume of 150 mL provided to the container. Further samples were taken on Days 4 and 5. Further feeding, at a feeding volume of 200 mL, was provided on Day 5. Further feeding was provided, and samples taken, on Day 6, at a feed volume of 400 mL. Samples were subsequently taken on Days 7 and 8.
[0534] Additionally, the following process, or specifically the following agitation, steps were used: [0535] 1. Days 0 to 3: none (i.e., static culture) [0536] 2. Days 3 to 4: rocking motion C (20 rpm, displacement amplitude of 20 mm) [0537] 3. Days 4 to 5: rocking motion B (20 rpm, displacement amplitude of 10 mm) [0538] 4. Days 6 to 8: linear compression (60 cpm, displacement amplitude of 20 mm)
[0539] The relevant agitation steps (either 1, 2, 3 or 4) are indicated in FIGS. 40 and 42 at the appropriate time intervals. The total viable cells are illustrated in FIGS. 40 and 42 for this particular agitation regime, whilst FIG. 42 also illustrates the cell viability.
[0540] The key outcomes of the primary T-cell run 2 were that: [0541] One vessel can used for all volumes: no manual transfers were required [0542] Rocking motion C (Day 3 to 4): cell death was observed [0543] Cell yield after 5 days of agitated mixing was >6?10.sup.6 [0544] Viability within release criteria throughout (?80%) [0545] As illustrated in FIG. 40, ?60% less culture medium used over 8 days in the present container compared to a standard WAVE perfusion protocol (estimated based on a standard protocol for perfusion culture of T lymphocytes in the WAVE Bioreactor System 2/10, GE Healthcare Life Sciences (Application note 28-9650-52 AC)).
[0546] It will be appreciated by persons skilled in the art that the above embodiment(s) have been described by way of example only and not in any limitative sense, and that various alterations and modifications are possible without departing from the scope of the disclosure as defined by the appended claims. Various modifications to the detailed designs as described above are possible, for example, variations may exist in shape, size, arrangement, assembly or the like.
[0547] In particular, any of the discussed actuation mechanisms may be utilized in any embodiment of the apparatuses, systems or methods discussed herein. Furthermore, any of the discussed holding elements may be utilized in any embodiment of the apparatuses, systems or methods discussed herein. Yet further, any of the discussed containers, platforms or other like cell processing components may be utilized in any embodiment of the apparatuses, systems of methods discussed herein.
[0548] Whilst the above examples also illustrate exemplary uses of the disclosed apparatus and system, the person skilled in the art would appreciate that it is equally applicable to other cell types, media types, transduction and activation reagents, and the like. Equally, the person skilled in the art would appreciate that other mixing/agitation regimes are equally contemplated as part of the disclosure, for example, the rate, the time periods, the amplitudes or the like of the rocking, swirling and/or compression regimes discussed herein.
