ROTATING CULTURE DEVICE AND CULTURE VESSEL FOR USE IN ROTATING CULTURE DEVICE

Abstract

The rotating culture device includes a tubular culture vessel long in an axial direction, a support mechanism which supports the culture vessel so that the culture vessel is rotatable about an axis thereof, rotation drive section which rotationally drives the culture vessel, tilting section which tilts the support mechanism so that the axis of the culture vessel is tilted in a vertical direction, and control section which controls operation of the rotation drive section and operation of the tilting section.

Claims

1. A rotating culture device comprising: a tubular culture vessel which is long in an axial direction; a support mechanism which supports the culture vessel so that the culture vessel is rotatable about an axis thereof; a rotation drive section which rotationally drives the culture vessel; a tilting section which tilts the support mechanism so that the axis of the culture vessel is tilted in a vertical direction; and a control section which controls operation of the rotation drive section and operation of the tilting section, wherein the culture vessel has a peripheral wall portion whose shape is cylindrical or a polygonal shape having eight or more sides so that a liquid culture medium in the culture vessel is prevented from being stirred to damage cells, and the control section controls the tilting section in accordance with a distribution state of cells so that uneven floating positions of the cells in the axial direction are dispersed in the axial direction.

2. A rotating culture device comprising: a tubular culture vessel which is long in an axial direction; a support mechanism which supports the culture vessel at both end sides thereof so that the culture vessel is rotatable about an axis thereof; a rotation drive section which rotationally drives the culture vessel; a reciprocating section which reciprocates the support mechanism in a horizontal direction along the axis of the culture vessel; and a control section which controls operation of the rotation drive section and operation of the reciprocating section, wherein the culture vessel has a peripheral wall portion whose shape is cylindrical or a polygonal shape having eight or more sides so that a liquid culture medium in the culture vessel is prevented from being stirred to damage cells, and the control section controls the reciprocating section in accordance with a distribution state of cells so that uneven floating positions of the cells in the axial direction are dispersed in the axial direction.

3. A rotating culture device comprising: a tubular culture vessel which is long in an axial direction; a support mechanism which supports the culture vessel at both end sides thereof so that the culture vessel is rotatable about an axis thereof; a rotation drive section which rotationally drives the culture vessel; a tilting section which tilts the support mechanism so that the axis of the culture vessel is tilted in a vertical direction; a reciprocating section which integrally reciprocates the support mechanism and the tilting section in a horizontal direction along the axis of the culture vessel; and a control section which controls operation of the rotation drive section, operation of the tilting section, and operation of the reciprocating section, wherein the culture vessel has a peripheral wall portion whose shape is cylindrical or a polygonal shape having eight or more sides so that a liquid culture medium in the culture vessel is prevented from being stirred to damage cells, and the control section controls the tilting section and the reciprocating section in accordance with a distribution state of cells so that uneven floating positions of the cells in the axial direction are dispersed in the axial direction.

4. The rotating culture device according to any one of claims 1 to 3, wherein the support mechanism includes a pair of support portions which rotatably support both end sides of the culture vessel, and the rotation drive section includes a drive mechanism which rotationally drives the culture vessel at one or both of the pair of support portions.

5. The rotating culture device according to claim 4, wherein each of the support portions includes a pair of support rollers which are parallel to the axis of the culture vessel and rotatably support an outer peripheral surface of an end portion of the culture vessel.

6. The rotating culture device according to claim 5, wherein the drive mechanism includes a mechanism which drives the pair of support rollers of the support portion to rotate in conjunction with each other.

7. The rotating culture device according to any one of claims 1 to 3, wherein the culture vessel has a peripheral wall composed of an observation window which allows an interior of the culture vessel to be seen therethrough, and an imaging section which captures the interior through the observation window is provided at an exterior position facing the peripheral wall.

8. The rotating culture device according to claim 7, wherein the culture vessel is provided, at an outer peripheral side of each of both end portions thereof, with a ring type illuminator which illuminates the interior through the observation window in coaxial relation to the culture vessel.

9. The rotating culture device according to claim 7, wherein the control section includes a position calculation unit which calculates positions of cultured cells captured by the imaging section, a storage unit which stores information on the positions of the cultured cells calculated by the position calculation unit, and a control unit which automatically controls the operation on the basis of the information on the positions stored in the storage unit.

10. The rotating culture device according to claim 7, wherein the culture vessel includes the peripheral wall portion that has a tubular shape and is composed of the observation window which allows the interior of the culture vessel to be seen therethrough, and end members which close both ends of the peripheral wall portion, each of the end members has an outer peripheral surface provided with a circumferential surface, the support mechanism includes a pair of support portions which rotatably support each of the end members of the culture vessel, and each of the support portions includes a pair of support rollers which are parallel to the axis of the culture vessel and rotatably support the circumferential surface of the end member.

11. The rotating culture device according to claim 7, wherein the culture vessel includes the peripheral wall portion that has a tubular shape and is composed of the observation window which allows the interior of the culture vessel to be seen therethrough, end members which close both ends of the peripheral wall portion, and an oxygen supply portion which extends from at least one of the end members along a central axis and is long in the axial direction, the oxygen supply portion includes a columnar core member and a gas permeable membrane attached over an outer peripheral surface of the core member, one or more helical recesses which communicate with an oxygen supply path formed within the end member are formed on the core member so as to extend in a manner of intersecting each other, and each of the helical recesses and the gas permeable membrane define a space therebetween, which serves as an oxygen flow passage through which oxygen flows, and the oxygen is supplied into the liquid culture medium through the gas permeable membrane.

12. The rotating culture device according to claim 10, wherein a cell suspension injection port, a cell taking-out port, a liquid culture medium supply port, and a liquid culture medium discharge port are provided in end surfaces of each of the end members.

13. A culture vessel for use in the rotating culture device according to any one of claims 1 to 3, wherein the culture vessel is formed in a tubular shape which is long in an axial direction, the culture vessel includes the peripheral wall portion that has a tubular shape and is composed of an observation window which allows an interior of the culture vessel to be seen therethrough, and end members which close both ends of the peripheral wall portion, and each of the end members has an outer peripheral surface including a circumferential surface.

14. The culture vessel according to claim 13, comprising an oxygen supply portion which extends from at least one of the end members along a central axis and is long in the axial direction, wherein the oxygen supply portion includes a columnar core member and a gas permeable membrane attached over an outer peripheral surface of the core member, one or more helical recesses which communicate with an oxygen supply path formed within the end member are formed on the core member so as to extend in a manner of intersecting each other, and each of the helical recesses and the gas permeable membrane define a space therebetween, which serves as an oxygen flow passage through which oxygen flows, and the oxygen is supplied into the liquid culture medium through the gas permeable membrane.

15. The culture vessel according to claim 13, wherein a cell suspension injection port, a cell taking-out port, a liquid culture medium supply port, and a liquid culture medium discharge port are provided in end surfaces of the end members.

16. The rotating culture device according to claim 11, wherein a cell suspension injection port, a cell taking-out port, a liquid culture medium supply port, and a liquid culture medium discharge port are provided in end surfaces of each of the end members.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0043] FIG. 1 is a perspective view showing a rotating culture device according to a representative embodiment of the present invention.

[0044] FIG. 2 is a longitudinal cross-sectional view of the rotating culture device taken along the axial direction of a culture vessel.

[0045] FIG. 3 is a plan view of the rotating culture device.

[0046] FIG. 4 is a perspective view showing the culture vessel attached to the rotating culture device.

[0047] FIG. 5 is a longitudinal cross-sectional view of the culture vessel.

[0048] FIG. 6A is a perspective view showing an oxygen supply portion of the culture vessel, and FIG. 6B is a transverse cross-sectional view of the oxygen supply portion.

[0049] FIG. 7 is a longitudinal cross-sectional view showing a main part of the culture vessel.

[0050] FIGS. 8A and 8B are explanatory diagrams showing a main part of a support mechanism of the rotating culture device that supports the culture vessel.

[0051] FIGS. 9A and 9B are explanatory diagrams showing a main part of the support mechanism.

[0052] FIG. 10 is a perspective view showing a main part of a rotation drive section of the rotating culture device that rotationally drives the culture vessel.

[0053] FIG. 11 is an exploded perspective view showing a main part of the rotating culture device.

[0054] FIG. 12 is a transverse cross-sectional view showing a main part of the rotating culture device.

[0055] FIG. 13 is an explanatory diagram showing a main part of the rotating culture device.

[0056] FIGS. 14A and 14B show explanatory diagrams for imagining a state where each operation is controlled.

[0057] FIGS. 15A to 15C show explanatory diagrams for imagining a state where each operation is controlled.

DESCRIPTION OF EMBODIMENTS

[0058] Next, an embodiment of the present invention will be described in detail with reference to the accompanying drawings.

[0059] As shown in FIGS. 1 to 3, a rotating culture device 1 of the present invention includes: a tubular culture vessel 10 which is long in the axial direction thereof; a support mechanism 11 which supports the culture vessel 10 at both end sides such that the culture vessel 10 is rotatable about an axis; rotation drive section 12 which rotationally drives the culture vessel; tilting section 13 which tilts the support mechanism 11 such that the axis of the culture vessel 10 is tilted in the vertical direction; reciprocating section 14 which integrally reciprocates the support mechanism 11 and the tilting section 13 in a horizontal direction along the axis of the culture vessel 10; an illuminator 17 which illuminates the interior of the culture vessel 10 through an observation window 10a provided at a peripheral wall portion 20 of the culture vessel 10; imaging section 16 which is provided at an outside position facing the peripheral wall portion 20 of the culture vessel 10 and captures an image of the interior of the culture vessel 10 through the observation window 10a; and control section 15 which controls operation of the rotation drive section 12, the tilting section 13, and the reciprocating section 14 on the basis of information of floating positions of cultured cells the image of which is captured by the imaging section 16.

[0060] Also as shown in FIGS. 4 and 5, the culture vessel 10 includes: the tubular peripheral wall portion 20 composed of the observation window 10a which allows the interior of the culture vessel 10 to be seen therethrough; end members 21A and 21B which close both ends of the peripheral wall portion 20; and an oxygen supply portion 10b which extends from at least one end member along the central axis and is long in the axial direction. As the material of the observation window, the material of an observation window used at an end surface of a conventional flat culture vessel can be widely used. In the case where the imaging section or the illuminator is omitted, the observation window can also be omitted. The shape of the peripheral wall portion 20 may be a shape other than a cylindrical shape, but is preferably a cylindrical shape or a shape close thereto such that an inside liquid culture medium is prevented from being stirred to damage cells. In the case where the shape of the peripheral wall portion 20 is a polygonal shape, the shape preferably has eight or more sides.

[0061] The end members 21A and 21B are provided with a cell suspension injection port 210, a cell taking-out port 211, a liquid culture medium supply port 212, and a liquid culture medium discharge port 213 which penetrate in the axial direction and are open in the end surfaces. In this embodiment, an example in which the cell suspension injection port 210 and the liquid culture medium supply port 212 are provided in the end member 21A, and the cell taking-out port 211 and the discharge port 213 are provided in the end member 21B is shown, but these ports are not limited to such a combination. For example, the four ports can be provided in one of the end members 21A and 21B, and no port can be provided in the other of the end members 21A and 21B.

[0062] By providing these ports in the end member 21A (21B), the necessity to provide these ports in the peripheral wall portion 20 is eliminated. Therefore, particularly in the case of a culture vessel which is long in the axial direction as in the present invention, the distribution of inside floating cells in the axial direction can be efficiently observed through the observation window 10a of the peripheral wall portion without being disturbed by these ports. In addition, regarding the configuration in which ports are provided in the end members as described above, the ports can be provided at any positions in the end surface of the end member 21A (21B), owing to a configuration in which the culture vessel 10 is supported by two support rollers 30A, 30A (30B, 30B) which rotatably support a circumferential surface 21a of the end member 21A (21B) as will be described later.

[0063] In this embodiment, a flow path of the oxygen supply portion 10b described later is open at the center positions of the end surfaces of the end members 21A and 21B such that oxygen can be supplied as appropriate during rotation of the culture vessel 10. Thus, the ports 210 to 213 are provided at positions displaced from the centers of the end surfaces, and work relating to injection of a cell suspension, taking-out of cells, and supply and discharge of a liquid culture medium through the respective ports is performed after the culture vessel 10 is detached from the device.

[0064] As a matter of course, when a rotation drive mechanism is configured such that these ports are not blocked, the above work can be performed with the culture vessel 10 attached to the device in a state where rotation is stopped. In addition, the oxygen supply portion 10b may be provided at a position displaced from the central axis, or the flow path of the oxygen supply portion 10b may be formed so as to be open only in one of the end members, whereby these ports may be located at the center of the end member such that the above work can be performed during rotation.

[0065] The circumferential surface 21a is provided in at least a part of the outer peripheral surface of each of the end members 21A and 21B. The circumferential surface 21a is a supported portion for supporting the culture vessel 10 by the support rollers 30A, 30A (30B, 30B) described later, and can be omitted in the case where another support mechanism is employed.

[0066] The size of the culture vessel 10 can be changed by changing the axial length or the diameter of the peripheral wall portion 20. By making the dimensions of the circumferential surfaces 21a of the end members 21A and 21B identical to each other even when the culture vessel 10 has any size, culture vessels 10 of various sizes can be employed without changing the structure of a support portion 11A (11B) which includes the support rollers 30A, 30A (30B, 30B).

[0067] Also as shown in FIGS. 6A and 6B, the oxygen supply portion 10b includes a columnar core member 22 and a gas permeable membrane 23 attached over the outer peripheral surface of the core member 22. One or more helical recesses 25 which communicate with an oxygen supply path 21b formed within the end member 21A and which extend such that the recesses 25 intersect each other are formed in the outer peripheral surface of the core member 22. Oxygen is supplied through gaps between the recesses 25 and the gas permeable membrane 23 as oxygen flow passages, and the supplied oxygen is supplied into a liquid culture medium substantially uniformly in the axial direction through the gas permeable membrane 23.

[0068] As the gas permeable membrane 23, a conventionally known one can be widely used. Reference character 24 denotes fixing rings each composed of an elastic member for fixing the gas permeable membrane 23, provided on the core member 22, at both end sides of the core member 22 and at positions outward of the oxygen flow passages.

[0069] As shown in FIG. 7, each helical recess 25 is formed so as to communicate with and join ring-shaped recessed grooves 26 formed at both end sides of the core member 22 in the circumferential direction. In both end portions 22a of the core member 22, recesses 220 open in the end surfaces and having bottoms are provided so as to be recessed in the axial direction at least to depths corresponding to the positions of the recessed grooves 26. In addition, one or more communication holes 260 are formed so as to extend from the bottom surface of each recessed groove 26 toward the inner peripheral surface of the recess 220.

[0070] Each end portion 22a of the core member 22 is inserted into an attachment recessed groove 27 formed so as to be open in the inner end surface of the end member 21A (21B), with a packing 28 interposed therebetween, and is fixed by: inserting an attachment bolt 29 having a through hole 290 serving as the oxygen supply path 21b into a communication hole 270 penetrating from the bottom surface of the recessed groove 27 to the outer end surface of the end member 21A (21B), from the outside in the axial direction; and threadedly engaging the attachment bolt 29 with a female screw groove formed in the recess 220 of the core member 22. The length of the attachment bolt 29 is set such that the communication holes 260 open in the inner surface of the recess 220 are not blocked.

[0071] Oxygen is gradually supplied into the liquid culture medium through a process in which oxygen is supplied through the through hole 290 of the attachment bolt 29 into the recess 220 of the core member 22, is supplied therefrom through the communication holes 260 into the gap between the gas permeable membrane 23 and the recessed groove 26 at the outer peripheral side, and then flows through a helical oxygen supply path between each recess 25 and the gas permeable membrane 23. In this embodiment, as shown in FIG. 2, oxygen is supplied from the support portion 11a side at which the end member 21A is supported, and oxygen is released through a gap between the end member 21B and the support portion 11B, which supports the end member 21B, into the atmosphere. In addition, oxygen may be forcedly supplied as in this embodiment, or the flow path may be merely open to the atmosphere.

[0072] In the oxygen supply portion 10b, the core member 22 is supported at both ends thereof by the end members 21A and 21B, but may be supported at one side thereof by one of the end members 21A and 21B. In addition, the core member 22 itself may be omitted, and oxygen may be supplied from the inner end surfaces of one or both of the end members 21A and 21B directly through the gas permeable membrane.

[0073] As shown in FIGS. 1 and 2, the support mechanism 11 includes: a flat-plate-shaped rotating support table 31 which is formed so as to be long along the axial direction of the culture vessel 10; and a pair of support portions 11A and 11B which are provided on the rotating support table 31 and support the end members 21A and 21B of the culture vessel 10 in a rotatable state. Also as shown in FIGS. 8A, 8B, 9A, and 9B, the support portion 11A (11B) includes the two support rollers 30A, 30A (30B, 30B) which rotatably support the circumferential surface 21a of the end member 21A (21B).

[0074] The two support rollers 30A, 30A (30B, 30B) each have an axis parallel to the axis of the culture vessel 10 and are disposed so as to be spaced apart from each other at an interval, and the circumferential surface 21a of the end member 21A (21B) is placed between these rollers and rotatably supported. Also as seen from FIG. 2, the support portion 11B having the support rollers 30B, 30B is attached via linear guides 34 provided on the rotating support table 31 along the longitudinal direction, that is, the axial direction of the supported culture vessel 10, such that the support portion 11B is movable in the axial direction along these guides.

[0075] As seen from FIG. 1, a fixing plate 40 having a long hole 400 which is long in the axial direction and through which a fixing screw 41 to be threadedly engaged with a screw hole of the support portion 11B is inserted is erected on a side end portion of the rotating support table 31. The long hole 400 is set with a predetermined length including at least a movable range along the linear guides 24 for the support portion 11B, and the position of the support portion 11B can be fixed by tightening the fixing screw 41 in a state where the support portion 11B has been moved in the axial direction. Accordingly, the position in the axial direction of the support portion 11B can be adjusted in accordance with the size of the culture vessel 10, particularly, the length in the axial direction of the culture vessel 10, and the end member of a culture vessel after size change can be similarly supported.

[0076] The support mechanism 11 of this embodiment allows the culture vessel 10 to be easily attached when the culture vessel 10 is merely placed thereon, and allows the culture vessel 10 to be easily detached, and an axis can be formed only on the basis of the accuracy of the circumferential surface 21a and the accuracy of the support rollers, so that axis alignment is not necessary and stable rotation is achieved at low cost. However, the present invention is not limited to such a support mechanism in any manner, and as a matter of course, a configuration in which one or both of the end members are supported so as to be rotatable about the axis can be employed.

[0077] Only one side may be supported so as to be rotatable about the axis and the other side may not be supported, or one side may be similarly supported as to be rotatable about the axis and an end member may be placed at the other side between two support rollers for rotatable support as in this embodiment. Alternatively, the peripheral wall portion 20 other than the end members can be placed between support rollers and supported.

[0078] As shown in FIGS. 1 and 2, as the rotation drive section 12, similar to the support mechanism 11, a drive mechanism 12A which rotationally drives the culture vessel 10 is provided on the flat-plate-shaped rotating support table 31 via the support portion 11A. More specifically, the drive mechanism 12A is a mechanism that rotationally drives the pair of support rollers 30A, 30A of the support portion 11A such that the support rollers 30A, 30A move in conjunction with each other, and the drive mechanism 12A is configured such that an endless belt 33 is entrained about pulleys 302, 302 provided at end portions of two shafts 301, 301 extending from the support rollers 30A, 30A and about a pulley 320 of a drive motor 32 as shown in FIG. 10 and the support rollers 30A, 30A are driven by the drive motor 32 in a state where the support rollers 30A, 30A move in conjunction with each other.

[0079] Accordingly, the support rollers 30A, 30A rotate in the same direction at the same speed, and the culture vessel 10 is stably rotationally driven through the circumferential surface 21a of the end member 21A. In this embodiment, the culture vessel 10 is rotationally driven through the support portion 11A at the fixed side at which no position adjustment mechanism is present. However, as a matter of course, the same rotation drive section 12 can be provided at each of the support portions 11A and 11B. In addition, both the support rollers 30A, 30A are driven, but a configuration in which only one of the support rollers 30A, 30A is driven can be employed.

[0080] On the rotating support table 31 on which the support mechanism 11 and the rotation drive section 12 which rotate the culture vessel 10 about the axis are provided, the imaging section 16 which captures an image of the interior of the culture vessel 10 and the illuminator 17 which illuminates the interior are further provided.

[0081] As shown in FIGS. 1 to 3, the imaging section 16 includes: a support frame 35 which is erected on a side end portion of the rotating support table 31; and an imaging box 37 in which an imaging camera 36 is housed and which is attached to the frame 35. The imaging camera 36 is set so as to face the center position of the culture vessel 10. In addition, the imaging box 37 in which the imaging camera 36 is housed is provided with a mechanism that can adjust the position of the imaging box 37 through the support frame 35 in the axial direction of the culture vessel 10 and in a horizontal direction that is perpendicular to the axis and is a direction toward/away from the culture vessel 10.

[0082] Specifically, the support frame 35 includes: a base plate 350 which is fixed to the side end portion of the rotating support table 31 and has a substantially L shape when being seen from the axial direction; an intermediate plate 351 which is horizontally attached to an upper end portion of the base plate 350 such that the position thereof in the axial direction can be changed; and an attachment plate 352 which similarly has a substantially L shape, is attached to the intermediate plate 351 such that the position thereof can be changed in a direction perpendicular to the axis, and has a front surface (a surface at the side facing the culture vessel 10) to which the imaging box 27 is attached. The position of the imaging box 37 relative to the culture vessel 10 can be adjusted by these two types of position adjustments, and a viewing angle can be set such that the full length of the interior of the culture vessel is included even when the size of the culture vessel 10, particularly, the length in the axial direction of the culture vessel 10, is changed, whereby setting can be made such that an image of the distribution state of floating cells in the axial direction can be captured at one time.

[0083] A plurality of screw holes 350b for threadedly engaging with attaching screws 38 penetrating communication holes 351a of the intermediate plate 351 are provided in the base plate 350 and in the axial direction such that the screw holes 350b can be selected. Accordingly, the position at which the intermediate plate 351 is attached can be adjusted in the axial direction. In addition, the communication holes 351a of the intermediate plate 351 are formed as long holes which are long in the axial direction, so that the position at which the intermediate plate 351 is attached can be finely adjusted in the axial direction after the screw holes 350b are selected. Moreover, the attachment plate 352 has communication holes 352a for inserting attaching screws 39 to be threadedly engaged with screw holes of the intermediate plate 351, and the communication holes 352a are formed as long holes which are long in a direction perpendicular to the axis, so that the position at which the attachment plate 352 is fixed can be adjusted in the direction perpendicular to the axis.

[0084] As shown in FIGS. 1 to 3 and 12, as the illuminator 17, a ring illuminator which illuminates the peripheral wall portion 20 of the culture vessel 10 is provided outside each of the end members 21A and 21B of the culture vessel 10. Support bases 170A and 170B of the respective ring illuminators 17 are attached via the linear guides 34, which are provided on the rotating support table 31, so as to be movable in the axial direction along these guides, similar to the support portion 11B.

[0085] The fixing plate 40 has long holes 401 and 402 which are long in the axial direction and through which fixing screws 42 and 43 to be threadedly engaged with screw holes of the respective support bases 170A and 170B are inserted, so that the position of the support base 170A/170B can be fixed by tightening the fixing screw 42/43 in a state where the support base 170A/170B has been moved in the axial direction. Accordingly, the position in the axial direction of the support base 170A/170B can be adjusted in accordance with the size of the culture vessel 10, particularly, the length in the axial direction of the culture vessel 10, and the support base 170A/170B can be similarly located outside the end member of the culture vessel after the change, whereby setting can be made such that the interior is illuminated substantially uniformly over the full length from both end sides.

[0086] As the color of the illumination of each illuminator 17, a color suitable for performing binarization processing for analysis on image data obtained by capturing an image of floating cells in the culture vessel with the imaging section 16 is selected in accordance with the type of the liquid culture medium. In addition, similarly, the position of each illuminator 17 is finely adjusted to a suitable position.

[0087] By the illuminators 17 illuminating the culture vessel 10 from both end sides of the culture vessel 10 as in this embodiment, the imaging section 16 is allowed to substantially uniformly capture an image of the entirety of the culture vessel 10. Other than using such ring illuminators, illuminators in various forms can be used, for example, one or more long bar-shaped illuminators which are parallel to the axis of the culture vessel 10 are provided at positions at which the illuminators do not obstruct image capturing by the imaging section 16. The illuminators 17 are used in such a manner that the illuminators 17 do not constantly illuminate the culture vessel but illuminate the culture vessel only for a predetermined short time period in synchronization with timing of capturing an image by the imaging section 16, in order to avoid influence of heat on cells.

[0088] As described above, the support mechanism 11 which supports the culture vessel 10 at both end sides such that the culture vessel 10 is rotatable about the axis, the rotation drive section 12 which rotationally drives the culture vessel, the illuminator 17 which illuminates the interior of the culture vessel 10, and the imaging section 16 which captures an image of the interior through the observation window 10a of the culture vessel 10 are provided on the rotating support table 31, and are integrally tilted or reciprocated together with the rotating support table 31 by the tilting section 13 or the reciprocating section 14.

[0089] Also as shown in FIGS. 1, 2, and 11, the tilting section 13 includes: a horizontal tilt support table 50 which is provided below the rotating support table 31; a support portion 51 which supports the rotating support table 31 at a substantially center portion in the longitudinal direction thereof such that the rotating support table 31 is rotatable about an axis in the width direction thereof; and an actuator 52 which drives one end side of the rotating support table 31 in the vertical direction to tilt the rotating support table 31 about the axis in the width direction, and the support portion 51 and the actuator 52 are provided on the tilt support table 50. A projection 44 having a through hole 440 which penetrates therethrough in the width direction is provided at a substantially center portion in the longitudinal direction of the lower surface of the rotating support table 31, and the support portion 51 includes: a support shaft 53 which is inserted through the through hole 440; and bearing portions 54 which are provided at both end portions of the tilt support table 50 and rotatably support both end portions 53a of the support shaft 53 projecting from the through hole 440.

[0090] The actuator 52 is a power cylinder such as a hydraulic, pneumatic, or electric cylinder in which a tube-side main body portion 520 is fixed at a head side thereof to a portion surrounding the opening of a hole 55 in the lower surface of the tilt support table 50, through which a rod 521 is passed, in a state where the rod 521 penetrates the hole 55 upward, and a distal end of the rod 521 is attached to an attachment portion 45 of the rotating support table 31 via a link 56 so as to be rotatable by a support pin 46. By the rod 521 of the actuator 52 moving in the vertical direction, the rotating support table 31 and the support mechanism 11, the rotation drive section 12, the illuminator 17, and the imaging section 16, which are fixed to the rotating support table 31, are integrally tilted relative to the tilt support table 50 about the support shaft 53, and the culture vessel 10, which is attached to the rotating support table 31, is also similarly tilted such that the axis thereof is tilted in the vertical direction.

[0091] As shown in FIGS. 11 and 13, proximity sensors 57 for detecting the position of a detection portion 47 fixed to the rotating support table 31 are provided to the tilt support table 50 and at a plurality of angular positions corresponding to the tilt range, in this embodiment, at an intermediate origin position and two limit positions. The control section 15 controls operation of the actuator 52 on the basis of signals of these sensors.

[0092] As shown in FIGS. 1, 2, and 11, the reciprocating section 14 includes: a horizontal reciprocation support table 60 which is provided further below the tilt support table 50 and has, at the lower surface side thereof, support legs 67 for placement on a floor or the like; a support portion 61 which supports the tilt support table 50 such that the tilt support table 50 is movable in the horizontal direction along the axis of the culture vessel which is the longitudinal direction; and an actuator 62 which moves the tilt support table 50 similarly in the horizontal direction along the axis of the culture vessel, and the support portion 61 and the actuator 62 are provided on the reciprocation support table 60.

[0093] As the support portion 61, linear guides 63, which movably support movable block portions 58 provided at appropriate locations on the lower surface of the tilt support table 50, at four corners in this embodiment, are provided on the upper surface of the reciprocation support table 60 and at positions corresponding to the movable block portions 58, respectively. The actuator 62 is a rocking motor such as a hydraulic or electric motor in which a main body portion 620 is fixed to a portion surrounding the opening of a hole 64 in the lower surface of the reciprocation support table 60, through which a rocking arm 621 is passed, in a state where the rocking arm 621 penetrates the hole 64 upward, and a distal end of the rocking arm 621 is attached to an attachment portion 59 of the tilt support table 50 via a link 65 so as to be rotatable by a support pin 590.

[0094] By the rocking arm 621 of the actuator 62 rocking, the rotating support table 31 and the support mechanism 11, the rotation drive section 12, the illuminator 17, and the imaging section 16, which are fixed to the rotating support table 31, are integrally linearly reciprocated relative to the reciprocation support table 60 along the horizontal linear guides 63 together with the tilt support table 50, and the culture vessel 10, which is attached on the rotating support table 31, is also horizontally and linearly reciprocated in the axial direction (in the direction of a line of projection to a horizontal plane when the culture vessel 10 is tilted).

[0095] As shown in FIGS. 1 and 11, proximity sensors 66 for detecting the position of a detection portion 500 fixed to the tilt support table 50 are provided to the reciprocation support table 60 and at a plurality of positions corresponding to the horizontal movement range, at a center position and two limit positions in this embodiment. The control section 15 controls operation of the actuator 62 on the basis of signals of these sensors.

[0096] Although not shown, the control section 15 is a computer which includes a calculation device and a storage device. The calculation device is mainly composed of a CPU such as a microprocessor, and various types of information are inputted thereto and outputted therefrom through an input-output portion or a bus line. The storage device is composed of a hard disk, a storage memory, or the like such as RAM and ROM inside or outside the calculation device, and programs specifying procedures of various processing operations by the calculation device, and processing data are stored therein.

[0097] The calculation device receives image data obtained by capturing an image of the interior of the culture vessel with the imaging section 16, stores the image data in the storage device, performs binarization processing on the image data to calculate the positions of cultured cells, and stores information on the positions of the cultured cells in the storage device. In addition, the calculation device determines operation amounts of the rotation drive section, the tilting section, and the reciprocating section by a predetermined program on the basis of the distribution of the position information of the cultured cells which is calculated and stored, and sends instructions.

[0098] Regarding the distribution of the position information of the cultured cells, a bias of the distribution can be calculated by using a known statistical calculation method from information such as a cell density in a certain region, and the respective operation amounts can be determined in accordance with the bias. Such instructions are automatically controlled so as to fall within a bias range which is preset by a feedback control method. That is, control is performed such that clumps of cells are scattered uniformly in the axial direction.

[0099] FIGS. 14A, 14B, 15A to 15C are explanatory diagrams for imagining a state where each operation is controlled by the control section 15. When inside, floating cells grow and the distribution thereof is biased below the axial position as shown in FIG. 14A, feedback control is performed such that the floating cells are distributed with the axial position as a center as shown in FIG. 14B, for example, by increasing the rotation speed of the culture vessel 10 by the rotation drive section. Regarding the control for vertical floating positions, control is also preferably performed using only image data or binarization data in a predetermined range in the axial direction, not using image data over the full length in the axial direction.

[0100] When the distribution of the inside floating cells becomes biased to one end side as shown in FIG. 15A, feedback control is performed such that the floating cells are distributed uniformly in the axial direction as shown in FIG. 15C by operating the tilting section and the reciprocating section, for example, to tilt the biased end side upward and horizontally move the entire culture vessel toward the other end side at the same time, as indicated by arrows in FIG. 15B, while the inside floating cells are maintained with good quality without violently colliding against each other. The tilting section and the reciprocating section can make various combinations of operations other than being operated in the same manner as described above, and as a matter of course, only one of the tilting section and the reciprocating section can be operated.

[0101] The control section 15 may be provided at any position. The control section 15 may be fixed to the rotating support table 31 or may be fixed to the reciprocation support table 60. Alternatively, the control section 15 may be connected at a distant position via a cable or the like. In addition, the method of control by the control section 15 is not limited to the above-described examples in any manner. The imaging section and the illuminator may be omitted, and tilt, reciprocation, and the like may be made periodically in a predetermined pattern. Alternatively, operation control may be performed manually on the basis of an imaging result.

[0102] Although the embodiment of the present invention has been described above, the present invention is not limited to such an embodiment in any manner. For example, the reciprocating section may be omitted and the culture vessel may be moved only with the rotation drive section and the tilting section, or the tilting section may be omitted and the culture vessel may be moved only with the rotation drive section and the reciprocating section. In addition, the tilting section and the support mechanism are integrally reciprocated in the embodiment, but the tilt support table and the reciprocation support table may be disposed such that the positions thereof are inverted vertically, and the reciprocating section and the support mechanism may be integrally tilted. Other than the above, as a matter of course, the present invention can be carried out in various modes without departing from the gist of the present invention.

DESCRIPTION OF THE REFERENCE CHARACTERS

[0103] 1 rotating culture device [0104] 10 culture vessel [0105] 10a observation window [0106] 10b oxygen supply portion [0107] 11 support mechanism [0108] 11A, 11B support portion [0109] 12 rotation drive section [0110] 12A drive mechanism [0111] 13 tilting section [0112] 14 reciprocating section [0113] 15 control section [0114] 16 imaging section [0115] 17 illuminator [0116] 20 peripheral wall portion [0117] 21A, 21B end member [0118] 21a circumferential surface [0119] 21b oxygen supply path [0120] 22 core member [0121] 22a end portion [0122] 23 gas permeable membrane [0123] 24 linear guide [0124] 25 recess [0125] 26 recessed groove [0126] 27 recessed groove [0127] 27 imaging box [0128] 28 packing [0129] 29 bolt [0130] 30A, 30B support roller [0131] 31 rotating support table [0132] 32 drive motor [0133] 33 endless belt [0134] 34 linear guide [0135] 35 support frame [0136] 36 imaging camera [0137] 37 imaging box [0138] 38 attaching screw [0139] 39 attaching screw [0140] 40 fixing plate [0141] 41, 42 fixing screw [0142] 44 projection [0143] 45 attachment portion [0144] 46 support pin [0145] 47 detection portion [0146] 50 tilt support table [0147] 51 support portion [0148] 52 actuator [0149] 53 support shaft [0150] 53a end portions [0151] 54 bearing portion [0152] 55 hole [0153] 56 link [0154] 57 proximity sensor [0155] 58 movable block portion [0156] 59 attachment portion [0157] 60 reciprocation support table [0158] 61 support portion [0159] 62 actuator [0160] 63 linear guide [0161] 64 hole [0162] 65 link [0163] 66 proximity sensor [0164] 67 support leg [0165] 70 cell [0166] 170A, 170B support base [0167] 210 injection port [0168] 211 taking-out port [0169] 212 supply port [0170] 213 discharge port [0171] 220 recess [0172] 260 communication hole [0173] 270 communication hole [0174] 290 through hole [0175] 301 shaft [0176] 302 pulley [0177] 320 pulley [0178] 350 base plate [0179] 350b screw hole [0180] 351 intermediate plate [0181] 351a communication hole [0182] 352 plate [0183] 352a communication hole [0184] 400 long hole [0185] 401 long hole [0186] 440 through hole [0187] 500 detection portion [0188] 520 main body portion [0189] 521 rod [0190] 590 support pin [0191] 620 main body portion [0192] 621 rocking arm