CELL CULTURE SYSTEM AND CELL CULTURE VESSEL
20170226465 · 2017-08-10
Assignee
Inventors
Cpc classification
C12M29/00
CHEMISTRY; METALLURGY
International classification
Abstract
A cell culture system of the present invention includes: a culture-medium storage part that holds a culture medium for culturing cells; a temporary holding part that is coupled to the culture-medium storage part and that is provided with a holding space for temporarily holding the culture medium supplied from the culture-medium storage part and a discharge port for discharging the culture medium; a culture-medium supply part that is coupled to the discharge port of the temporary holding part and that supplies the culture medium discharged from the temporary holding part, to a cell culture vessel; and a discharge part that discharges the culture medium from the cell culture vessel, wherein the culture-medium supply part is provided with a culture-medium intermittent supply mechanism that intermittently supplies the culture medium discharged from the temporary holding part, to the cell culture vessel in a predetermined cycle.
Claims
1. A cell culture system comprising: a culture-medium storage part that holds a culture medium for culturing cells; a temporary holding part that is coupled to the culture-medium storage part and that is provided with a holding space for temporarily holding the culture medium supplied from the culture-medium storage part and a discharge port for discharging the culture medium; a culture-medium supply part that is coupled to the discharge port of the temporary holding part and that supplies the culture medium discharged from the temporary holding part, to a cell culture vessel; and a discharge part that discharges the culture medium from the cell culture vessel, wherein the cell culture vessel is coupled to the culture-medium supply part at a position lower in the direction of gravitational force than the culture-medium supply part; and the culture-medium supply part is provided with a flow passage that allows the culture medium to be intermittently supplied from the temporary holding part to the cell culture vessel according to the siphon principle.
2. A cell culture system comprising: a culture-medium storage part that holds a culture medium for culturing cells; a temporary holding part that is coupled to the culture-medium storage part and that is provided with a holding space for temporarily holding the culture medium supplied from the culture-medium storage part and a discharge port for discharging the culture medium; a culture-medium supply part that is coupled to the discharge port of the temporary holding part and that supplies the culture medium discharged from the temporary holding part, to a cell culture vessel; and a discharge part that discharges the culture medium from the cell culture vessel, wherein the cell culture vessel is coupled to the culture-medium supply part at a position lower in the direction of gravitational force than the culture-medium supply part; and the culture-medium supply part is provided with a supply gate that opens a flow passage when the culture medium in the holding space reaches a predetermined amount.
3. A cell culture system according to claim 1, wherein the culture-medium supply part is provided with the flow passage that causes the culture medium before being supplied to the cell culture vessel to pass through a position that is higher in the direction of gravitational force than the discharge port of the temporary holding part and that is lower in the direction of gravitational force than the maximum height of the holding space.
4. A cell culture system according to claim 3, wherein the discharge port is disposed in a side surface of the temporary holding part at the position of a predetermined height from a bottom surface of the temporary holding part; and the culture-medium supply part forms the flow passage from the discharge port to the outside of the temporary holding part.
5. A cell culture system according to claim 3, wherein the culture-medium supply part forms the flow passage from the discharge port of the temporary holding part to the inside of the temporary holding part; and an end of the culture-medium supply part that is opposite from an end thereof that is connected to the discharge port of the temporary holding part has an opening in the vicinity of the bottom surface of the temporary holding part with a gap therebetween.
6. A cell culture system according to claim 2, wherein the supply gate opens the flow passage when the culture medium in the holding space reaches a predetermined weight.
7. A cell culture system according to claim 2, wherein the supply gate opens the flow passage when the top surface of the culture medium in the holding space reaches a predetermined height.
8. A cell culture system according to claim 1, wherein the cell culture vessel is provided with a discharge port for discharging the culture medium to the outside when the height of the top surface of the culture medium therein reaches a predetermined height.
9. A cell culture system according to claim 1, further comprising a supply-rate adjusting part that controls the supply rate of the culture medium supplied from the culture-medium storage part to the temporary holding part.
10. A cell culture vessel comprising: a supply port for supplying a culture medium; a discharge port for discharging the culture medium; and a tubular member that is connected to the discharge port, wherein the tubular member forms a flow passage that allows the culture medium in the cell culture vessel to be intermittently discharged according to the siphon principle.
11. A cell culture vessel according to claim 10, wherein the tubular member forms a flow passage that passes through a position that is higher in the direction of gravitational force than the discharge port and that is lower in the direction of gravitational force than the maximum height of a space in the cell culture vessel.
12. A cell culture vessel according to claim 11, wherein the discharge port is disposed in a side surface of the cell culture vessel at the position of a predetermined height from the bottom surface of the cell culture vessel; and the tubular member forms a flow passage from the discharge port to the outside of the cell culture vessel.
13. A cell culture vessel according to claim 11, wherein the tubular member forms a flow passage from the discharge port to the inside of the cell culture vessel; and an end of the tubular member that is opposite from an end thereof that is connected to the discharge port has an opening in the vicinity of a bottom surface of the cell culture vessel with a gap therebetween.
14. A cell culture system comprising: a cell culture vessel according to claim 10; and a culture-medium supply part that supplies the culture medium from the supply port of the cell culture vessel to the inside thereof.
15. A cell culture system according to claim 2, wherein the cell culture vessel is provided with a discharge port for discharging the culture medium to the outside when the height of the top surface of the culture medium therein reaches a predetermined height.
16. A cell culture system according to claim 2, further comprising a supply-rate adjusting part that controls the supply rate of the culture medium supplied from the culture-medium storage part to the temporary holding part.
Description
BRIEF DESCRIPTION OF DRAWINGS
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DESCRIPTION OF EMBODIMENTS
[0060] Cell culture systems according to embodiments of the present invention will be described below with reference to the drawings.
First Embodiment
[0061] A cell culture system 100 of this embodiment is a system that has a configuration shown in
[0062] A culture-medium storage means (culture-medium storage part) 3 is installed outside an incubator 1 and stores a culture medium (cell culture solution) therein. A temperature control means (temperature control part) 2 is provided in order to maintain the temperature of the culture medium at an appropriate temperature for storage (for example, 4° C.).
[0063] The culture-medium storage means 3 is coupled to a temporary holding means (temporary holding part) 4 that is installed in the incubator 1, via a tubular member (tube or the like). Because the culture-medium storage means 3 is installed at a position higher in the direction of gravitational force than the temporary holding means 4, the culture medium in the culture-medium storage means 3 is gravitationally supplied to the temporary holding means 4 via the tubular member (tube or the like). The culture medium supplied to the temporary holding means 4 is held in a space (holding space) in the temporary holding means 4 and is heated to a temperature in the incubator (for example, 37° C.) that is an appropriate temperature for cell culturing.
[0064] The tubular member that connects the culture-medium storage means 3 and the temporary holding means 4 is provided with a supply-rate adjusting means (supply-rate adjusting part) 5, so that the flow rate of the culture medium flowing in the tubular member is controlled, thereby making it possible to adjust the supply rate of the culture medium from the culture-medium storage means 3 to the temporary holding means 4.
[0065] The culture medium heated, in the temporary holding means 4, to an appropriate temperature for cell culturing (for example, 37° C.) is supplied, in the incubator 1, to cell culture vessels 7 via a culture-medium supply means (culture-medium supply part) 6. The temporary holding means 4 is provided with: a supply port 4a through which the culture medium is supplied from the culture-medium storage means 3 via the tubular member; and a discharge port 4b through which the culture medium is discharged to the culture-medium supply means 6. Although the position where the supply port 4a is installed is arbitrarily decided, it is preferred to install the supply port 4a at an upper portion of the temporary holding means 4 to cause the supplied culture medium to be dripped into the inner space. Accordingly, backflow of the culture medium can be prevented, thereby making it possible to prevent the culture medium in the culture-medium storage means 3 from being contaminated (the occurrence of contamination). On the other hand, it is preferred that the installation site of the discharge port 4b be a place allowing the culture medium in the temporary holding means 4 to be completely discharged, i.e., a bottom surface or a lower portion on a side surface of the temporary holding means 4.
[0066] The culture-medium supply means 6 is provided with a siphon mechanism 6a (culture-medium intermittent supply mechanism). The siphon mechanism 6a is connected to the discharge port 4b of the temporary holding means 4 and has a structure in which a flow passage thereof is turned around, as shown in
[0067] The flow passage of the siphon mechanism 6a extends upward in the direction of gravitational force from the discharge port 4b of the temporary holding means 4 to the turn-around point 6b, passes through the turn-around point 6b, extends downward in the direction of gravitational force to reach a branching means (branching part) 6c. From the branching means 6c, the flow passage is split into a plurality of tubular members (tubes or the like). The plurality of split tubular members are respectively connected to separate cell culture vessels 7 to supply the culture medium thereto.
[0068] The cell culture vessels 7 each have a supply port 7a to which the tubular member of the culture-medium supply means 6 is connected and a discharge port 7b through which the culture medium is discharged to the outside of the cell culture vessel. The discharge port 7b is disposed in a side surface of the cell culture vessel 7, and, when the amount of the culture medium (the height of the culture medium) in the cell culture vessel exceeds a certain value, the culture medium is discharged to a culture-medium discharge means (discharge part) 8 through the discharge port 7b. The amount of the culture medium held in the cell culture vessel 7 is determined by the installation height of the discharge port 7b. On the other hand, although the position where the supply port 7a is installed is arbitrarily decided, when it is installed as far away from the discharge port 7b as possible, the replacement efficiency of the culture medium can be increased. Furthermore, when the supply port 7a is installed at an upper portion of the cell culture vessel 7 to cause the supplied culture medium to be dripped into the cell culture vessel, backflow of the culture medium can be prevented, thus making it possible to prevent the culture medium in the culture-medium supply means 6, the temporary holding means 4, and the culture-medium storage means 3, which are disposed at upstream sides of the system, from being contaminated (the occurrence of contamination).
[0069] The culture-medium discharge means 8 is provided with a waste-liquid holding means 8a, and the waste-liquid holding means 8a has: waste-liquid supply ports 8b to which tubular members extending from the cell culture vessels 7 are connected; and a waste-liquid discharge port 8c through which the culture medium is discharged to the outside of the waste-liquid holding means 8a. The waste-liquid supply ports 8b are installed in a top surface of the waste-liquid holding means 8a, and the culture medium supplied through each of the waste-liquid supply ports 8b is dripped into a space in the waste-liquid holding means 8a and is discharged through the waste-liquid discharge port 8c. In this way, when the culture medium is dripped into the space in the waste-liquid holding means 8a, backflow of the culture medium can be prevented, thus making it possible to prevent the insides of the cell culture vessels 7 from being contaminated (the occurrence of contamination).
[0070] Next, the supply-rate adjusting means 5 will be described.
[0071] The supply-rate adjusting means 5 is disposed in the tubular member (tube or the like) that connects the discharge port 3a of the culture-medium storage means 3 and the supply port 4a of the temporary holding means 4, applies an external force to deform the tubular member, thus reducing the cross-section area of a tubular-member lumen, thereby limiting the flow volume of the solution to suppress the flow rate. On the contrary, when the external force is released, the tubular member recovers to the original state due to the elastic force of the tubular member, thus making it possible to increase the flow rate. In this way, the supply-rate adjusting means 5 adjusts the flow rate of the solution flowing in the tubular member due to the magnitude of the external force applied to the tubular member.
[0072] Furthermore, a liquid feeding pump, such as a peristaltic pump, may be used as the supply-rate adjusting means 5. In this case, the culture-medium storage means 3 need not be disposed at a position higher than the temporary holding means 4 in the direction of gravitational force, thus increasing the degree of freedom in the installation site of the culture-medium storage means 3.
[0073] Next, an example procedure for replacing the culture medium by using the culture system 100 of this embodiment will be described.
[0074] A user of this system first sets the system in a state in which the supply rate of the culture medium is zero, i.e., in a state in which supply of the culture medium is stopped, by using the supply-rate adjusting means 5 and replenishes the culture-medium storage means 3 with the culture medium. The user prepares the cell culture vessels 7 in which the culture medium and cells are contained, connects, in the incubator, the supply ports 7a of the cell culture vessels 7 to the tubular members of the culture-medium supply means 6, and connects the discharge ports 7b of the cell culture vessels 7 to the waste-liquid supply ports 8b of the waste-liquid holding means 8a via the tubular members.
[0075] When culture-medium replacement is needed, the user first adjusts the supply-rate adjusting means 5 and sets the supply rate to an appropriate rate. When the state in which the supply rate is zero is released by means of the supply-rate adjusting means 5, the culture medium starts dripping into the temporary holding means 4 due to the force of gravity. In adjusting the supply rate, the supply rate may be set to a rate determined in advance or may be adjusted to an appropriate supply rate while the user checks visually.
[0076] The culture medium is supplied to the temporary holding means 4, and the culture medium level in the temporary holding means 4 rises (
[0077] The culture medium discharged from the temporary holding means 4 is supplied to the plurality of cell culture vessels 7 via the branching means 6c of the culture-medium supply means 6. When exceeding a specified amount in the cell culture vessels 7, the culture medium is discharged from the discharge ports 7b of the cell culture vessels 7 and is discharged to the outside of the incubator via the waste-liquid holding means 8a. Accordingly, the old culture medium in the cell culture vessels 7 is replaced with a new culture medium, thereby making it possible to reduce the rate of deterioration of the culture medium.
[0078] In this embodiment, as the siphon mechanism 6a, a tubular member (tube or the like) may form a flow passage, as shown in
Second Embodiment
[0079] A cell culture system 300 of this embodiment is a system that has a configuration shown in
[0080] The culture-medium storage means 3 is installed outside the incubator 1 and stores the culture medium (cell culture solution) therein. The temperature control means 2 is provided in order to maintain the temperature of the culture medium to an appropriate temperature for storage (for example, 4° C.).
[0081] The culture-medium storage means 3 is coupled to the temporary holding means 4, which is installed in the incubator via the tubular member (tube or the like). Because the culture-medium storage means 3 is installed at a position higher in the direction of gravitational force than the temporary holding means 4, the culture medium in the culture-medium storage means 3 is gravitationally supplied to the temporary holding means 4 via the tubular member (tube or the like). The culture medium supplied to the temporary holding means 4 is heated to a temperature in the incubator (for example, 37° C.) that is an appropriate temperature for cell culturing.
[0082] The tubular member that connects the culture-medium storage means 3 and the temporary holding means 4 is provided with the supply-rate adjusting means 5, and the flow rate of the culture medium flowing in the tubular member is controlled, thereby making it possible to adjust the supply rate of the culture medium from the culture-medium storage means 3 to the temporary holding means 4.
[0083] The culture medium heated, in the temporary holding means 4, to an appropriate temperature for cell culturing (for example, 37° C.) is supplied, in the incubator 1, to the cell culture vessels 7 via the culture-medium supply means 6. The temporary holding means 4 is provided with: the supply port 4a, through which the culture medium is supplied from the culture-medium storage means 3 via the tubular member; and the discharge port 4b, through which the culture medium is discharged to the culture-medium supply means 6. Although the position where the supply port 4a is installed is arbitrarily decided, it is preferred to install the supply port 4a at an upper portion of the temporary holding means 4 to cause the supplied culture medium to be dripped into the inner space. Accordingly, backflow of the culture medium can be prevented, thereby making it possible to prevent the culture medium in the culture-medium storage means 3 from being contaminated (the occurrence of contamination). On the other hand, it is preferred that the installation site of the discharge port 4b be a place in the temporary holding means 4 for allowing the culture medium therein to be completely discharged.
[0084] The culture-medium supply means 6 is provided with a supply gate 6d (culture-medium intermittent supply mechanism) at a connection part with the temporary holding means 4. The supply gate 6d is a means that is capable of opening and closing the flow passage according to the amount of the culture medium in the temporary holding means 4, that opens the gate to discharge the culture medium from the temporary holding means 4 when the amount of the culture medium in the temporary holding means 4 exceeds a predetermined amount, and that closes the gate to block discharge of the culture medium from the temporary holding means 4 when the culture medium in the temporary holding means 4 decreases to the predetermined amount. The supply gate 6d is connected to the cell culture vessels 7 via the tubular members.
[0085] When reaching the branching means 6c, the tubular member of the culture-medium supply means 6 extending from the supply gate 6d is split into a plurality of tubular members. The plurality of split tubular members are respectively connected to the separate cell culture vessels 7 to supply the culture medium thereto.
[0086] The cell culture vessels 7 each have the supply port 7a, to which the tubular member of the culture-medium supply means 6 is connected, and the discharge port 7b, through which the culture medium is discharged to the outside of the cell culture vessel. The discharge port 7b is disposed in a side surface of the cell culture vessel 7, and, when the amount of the culture medium (the height of the culture medium) in the cell culture vessel exceeds a certain value, the culture medium is discharged to the culture-medium discharge means 8 via the discharge port 7b. The amount of the culture medium held in the cell culture vessel is determined by the installation height of the discharge port 7b. On the other hand, although the position where the supply port 7a is installed is arbitrarily decided, when it is installed as far away from the discharge port 7b as possible, the replacement efficiency of the culture medium can be increased. Furthermore, when the supply port 7a is installed at an upper portion of the cell culture vessel 7 to cause the supplied culture medium to be dripped into the cell culture vessel 7, backflow of the culture medium can be prevented, thus making it possible to prevent the culture medium in the culture-medium supply means 6, the temporary holding means 4, and the culture-medium storage means 3, which are disposed at upstream sides of the system, from being contaminated (the occurrence of contamination).
[0087] The culture-medium discharge means 8 is provided with the waste-liquid holding means 8a, and the waste-liquid holding means 8a has: the waste-liquid supply ports 8b, to which the tubular members extending from the cell culture vessels 7 are connected; and the waste-liquid discharge port 8c, through which the culture medium is discharged to the outside of the waste-liquid holding means 8a, The waste-liquid supply ports 8b are installed in the top surface of the waste-liquid holding means 8a, and the culture medium supplied through each of the waste-liquid supply ports 8b is dripped into a space in the waste-liquid holding means 8a and is discharged through the waste-liquid discharge port 8c. In this way, when the culture medium is dripped into the space in the waste-liquid holding means 8a, backflow of the culture medium can be prevented, thus making it possible to prevent the insides of the cell culture vessels 7 from being contaminated (the occurrence of contamination).
[0088] The supply-rate adjusting means 5 is the same as that in the first embodiment.
[0089] Next, the supply gate 6d will be described. The supply gate 6d is a means capable of opening and closing the flow passage according to the amount of the culture medium in the temporary holding means 4 and has a configuration shown in
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[0092] Next, an example procedure for replacing the culture medium by using the culture-medium replacement system 300 of this embodiment will be described.
[0093] A user of this system first sets the system in a state in which the supply rate of the culture medium is zero, i.e., in a state in which supply of the culture medium is stopped, by using the supply-rate adjusting means 5 and replenishes the culture-medium storage means 3 with the culture medium. The user prepares the cell culture vessels 7 in which the culture medium and cells are contained, connects, in the incubator, the supply ports 7a of the cell culture vessels 7 to the tubular members of the culture-medium supply means 6, and connects the discharge ports 7b of the cell culture vessels 7 to the waste-liquid supply ports 8b of the waste-liquid holding means 8a via the tubular members.
[0094] When culture-medium replacement is needed, the user first adjusts the supply-rate adjusting means 5 and sets the supply rate to an appropriate rate. When the state in which the supply rate is zero is released by means of the supply-rate adjusting means 5, the culture medium starts dripping into the temporary holding means 4 due to the force of gravity. In adjusting the supply rate, the supply rate may be set to a rate determined in advance or may be adjusted to an appropriate supply rate while the user checks visually.
[0095] The culture medium is supplied to the temporary holding means 4, and the amount of the culture medium in the temporary holding means 4 increases. When the pressure from the culture medium in the temporary holding means 4 on the supply gate 6d exceeds the threshold, the gate opens, and the culture medium in the temporary holding means 4 starts to be discharged via the culture-medium supply means 6, keeps being discharged until the pressure from the culture medium in the temporary holding means 4 on the supply gate 6d decreases to the threshold, and stops being discharged when the pressure decreases to the threshold. Then, when the amount of the culture medium in the temporary holding means 4 starts to increase due to the culture medium supplied from the culture-medium storage means 3, and the pressure from the culture medium in the temporary holding means 4 on the supply gate 6d exceeds the threshold, discharge occurs again. This process is repeated.
[0096] The culture medium discharged from the temporary holding means 4 is supplied to the plurality of cell culture vessels 7 via the branching means 6c of the culture-medium supply means 6. When exceeding a specified amount in the cell culture vessels 7, the culture medium is discharged from the discharge ports 7b of the cell culture vessels 7 and is discharged to the outside of the incubator via the waste-liquid holding means 8a. Accordingly, the old culture medium in the cell culture vessels 7 is replaced with a new culture medium, thereby making it possible to reduce the rate of deterioration of the culture medium.
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Third Embodiment
[0100] A cell culture system 500 of this embodiment differs from those of the above-described embodiments in that a pressure applying means 51 that applies a pressure to the culture medium in the culture-medium storage means 3 is provided, as shown in
[0101] As the pressure applying means 51, a means for pumping gas into the culture-medium storage means 3 by means of a pump can be used, for example. Accordingly, the pressure of an air space in the culture-medium storage means 3 is increased, thus making it possible to apply an external pressure to the culture medium. It is preferred that gas to be pumped into the culture-medium storage means 3 be sterilized. As shown in
[0102] Furthermore, it is also possible to adopt a form in which the culture-medium holding means has a column-shaped structure, the isolation member has a disk-shaped structure that fits in the inner wall of the column-shaped structure while maintaining airtightness, and the inner wall of the column-shaped structure and the circumference of the disk-shaped structure have screw structures to be engaged with each other. In this case, when the isolation member is attached to the inside of the culture-medium holding means and is rotated in the disk circumferential direction, the isolation member is moved in the height direction of the column structure, thus making it possible to apply a pressure to the culture medium in the culture-medium holding means (or remove the pressure therefrom). The rotation of the isolation member may be mechanically performed, and the rotation of the isolation member may be controlled by the control unit in a wired or wireless manner.
[0103] Here, it is preferred that the pressure applying means 51 be controlled such that the pressure in the culture-medium storage means 3 does not increase to a certain level or more.
[0104] Note that, in this embodiment, the culture-medium storage means 3 is not necessarily disposed at a higher position than the temporary holding means 4. Furthermore, the temporary holding means 4 is not necessarily disposed at a higher position than the culture-medium supply means 6.
Fourth Embodiment
[0105] As shown in
[0106] As the negative-pressure supply means 71, it is possible to adopt a means that is provided with a pump 81 and a waste-liquid container 82 shown in
[0107] As the pump of the negative-pressure supply means, a liquid feeding pump, such as a peristaltic pump, may be used. In this case, the liquid feeding pump may be disposed on a tubular member of the suction port 83.
Fifth Embodiment
[0108] As shown in
[0109] In this embodiment, the supply-rate adjusting means 5 can exchange information with the control unit 19, which is provided outside the incubator, in a wired or wireless manner, and the flow rate of a solution, such as a culture medium, flowing in the tubular member (tube or the like) can be remotely controlled.
[0110] When culture-medium replacement is needed, the user remotely adjusts the supply-rate adjusting means 5 by using the control unit 19 and sets the supply rate to an appropriate rate. In adjusting the supply rate, the supply rate may be set to a rate determined in advance or may be adjusted to an appropriate supply rate while monitoring a drip rate by using a monitoring system (not shown).
[0111] According to this embodiment, the user can start to remotely replace the culture medium or can remotely change the supply rate, at arbitrary timing during the cell culturing. For example, when used with a system (not shown) capable of remotely monitoring the state of the cells, it is possible to start to remotely replace the culture medium or remotely change the supply rate at arbitrary timing during the cell culturing in accordance with the state of the cells. By doing so, because the user can work without entering a work space, it is possible to save the trouble of having to change into disposable work clothes and the costs thereof and to reduce the risk of contamination of the cell culture system by bacteria etc.
[0112] The control unit 19 of this embodiment may be able to exchange information with the temperature control means 2, the pressure applying means 51, and the negative-pressure supply means 71, which are provided in the above-described embodiments, in a wired or wireless manner and may be able to remotely control them. Accordingly, it is possible to improve the user's remote work efficiency.
[0113] In the above-described embodiments, a description has been given of a case in which the discharge port of each cell culture vessel has a form shown in
[0114]
[0115] Then, the culture medium is supplied from the culture-medium supply means 6, and, when the height of the culture medium in the cell culture vessel reaches a position higher than the turn-around point 7e of the flow passage of the discharge siphon mechanism 7d, discharge occurs again. This process is repeated. Here, the discharge siphon mechanism 7d functions as a culture-medium intermittent discharge mechanism.
[0116]
[0117] Then, the culture medium is supplied from the culture-medium supply means 6, and, when the height of the culture medium in the cell culture vessel reaches the predetermined height (7f), discharge occurs again. This process is repeated. The predetermined height (7f) is set according to the type of cells to be cultured and the culture conditions.
[0118] Although
[0119] In the above-described embodiments, although the flask-like culture vessels are shown in the figures as the cell culture vessels to be used, it is also possible to use petri-dish-like culture vessels, as shown in
[0120] In the forms that are provided with the siphon mechanisms of the above-described embodiments, although a case in which a single siphon mechanism of the culture-medium supply means is provided has been illustrated, it is also possible to provide a plurality of siphon mechanisms, as shown in
[0121] In the above-described embodiments, as shown in
[0122] The above-described embodiments show a form in which the culture-medium storage means 3 is coupled to the temporary holding means 4 via the tubular member (tube or the like), the culture-medium storage means 3 is disposed at a position higher in the direction of gravitational force than the temporary holding means 4, and thus, the culture medium in the culture-medium storage means 3 is gravitationally supplied to the temporary holding means 4 via the tubular member (tube or the like); however, for example, as shown in
[0123] In the above-described embodiments, for example, as shown in
[0124] In the above-described embodiments, an example case in which a waste liquid discharged through the discharge ports of the culture vessels is discharged via the waste-liquid holding means has been illustrated; however, for example, as shown in
[0125] In this case, in the third embodiment, the suction port of the negative-pressure supply means may be connected to the discharge ports of the cell culture vessels.
[0126] In the above-described embodiments, although a form in which a single culture-medium storage means and a single temporary holding means are provided has been illustrated, a plurality of culture-medium storage means and a plurality of temporary holding means may be provided. In this case, multiple types of culture media can be supplied to the cell culture vessels.
[0127] In the above-described embodiments, although a form in which connections are made from a single culture-medium supply means to a plurality of cell culture vessels has been illustrated, it is also possible to adopt a form in which a connection may be made from a single culture-medium supply means to a single cell culture vessel.
[0128] In the above-described embodiments, a culture-medium temperature monitoring means that monitors the culture-medium temperature may be provided in the temporary holding means. At this time, information of the culture-medium temperature monitored by the culture-medium temperature monitoring means may be remotely sent to the control unit. Thus, it is possible to prevent the culture medium at a temperature that is unsuitable for cell culturing from being accidentally supplied.
[0129] Furthermore, the amount of the culture medium held in each cell culture vessel is set to the lowest possible amount, thereby making it possible to improve the culture-medium replacement efficiency. The amount of the culture medium held in the cell culture vessel may be optimized depending on the type of cells to be cultured.
[0130] In the above-described embodiments, the control unit, which is provided outside the incubator, may be able to exchange information in a wired or wireless manner with the temperature control means, the pressure applying means, the negative-pressure supply means, and the liquid feeding pump, which are provided in the above-described embodiments, and may be able to remotely control them. Accordingly, it is possible to improve the user's remote work efficiency.
[0131] In the above-described embodiments, in the form in which the user remotely controls culture-medium replacement by using the control unit, the control unit may remotely control culture-medium replacement on the basis of a schedule (program) set in advance by the user.
[0132] In the above-described embodiments, although a form in which the temporary holding means is disposed inside the incubator has been illustrated, the temporary holding means may be disposed outside the incubator. In that case, it is preferred to provide a temperature control means for maintaining the solution in the temporary holding means at an appropriate temperature for cell culturing (for example, 37° C.).
[0133] Furthermore, in the above-described embodiments, although a form in which the waste-liquid holding means is disposed outside the incubator has been illustrated, for example, as shown in
[0134] A PC can be shown as an example of the control unit of the present invention, and the PC can perform control performed by the control unit in the above-described embodiments.
[0135] Specifically, the control unit is, for example, a PC that has a CPU and a memory, and, the CPU executes a control program written in the memory, thereby realizing the functions as the control unit.
[0136] In the present invention, the temperature of the culture medium in the culture-medium storage means 3 may be set at an appropriate temperature according to the culture conditions. The culture-medium temperature may be maintained at 37° C. by means of the temperature control means 2. Furthermore, the culture-medium storage means 3 may be disposed inside the incubator, and, in that case, if the culture-medium temperature is maintained at 37° C., the temperature control means 2 may be omitted.
[0137] In the present invention, the culture-medium storage means and the temporary culture-medium holding means are means capable of holding the culture medium or another solution (for example, a cleaning solution or the like) and serve as solution holding means. Each of the culture-medium storage means and the temporary culture-medium holding means serves as a solution holding means, and both of the means, as a unit, also serve as a solution holding means.
[0138] The present invention can provide a cell culture system including:
[0139] a culture-medium storage means that holds a culture medium for culturing cells;
[0140] a temporary holding means that is coupled to the culture-medium storage means and that is provided with a holding space for temporarily holding the culture medium supplied from the culture-medium storage means and a discharge port for discharging the culture medium;
[0141] a culture-medium supply means that is coupled to the discharge port of the temporary holding means and that supplies the culture medium discharged from the temporary holding means, to a cell culture vessel; and
[0142] a culture-medium discharge means that discharges the culture medium from the cell culture vessel,
[0143] wherein the culture-medium supply means is provided with a culture-medium intermittent supply mechanism that intermittently supplies the culture medium discharged from the temporary holding means, to the cell culture vessel in a predetermined cycle.
[0144] The present invention can provide a cell culture system including:
[0145] a culture-medium storage means that holds a culture medium for culturing cells;
[0146] a temporary holding means that is coupled to the culture-medium storage means and that is provided with a holding space for temporarily holding the culture medium supplied from the culture-medium storage means and a discharge port for discharging the culture medium;
[0147] a culture-medium supply means that is coupled to the discharge port of the temporary holding means and that supplies the culture medium discharged from the temporary holding means, to a cell culture vessel; and
[0148] a culture-medium discharge means that discharges the culture medium from the cell culture vessel,
[0149] wherein the culture-medium discharge means is provided with a culture-medium intermittent discharge mechanism that intermittently discharges the culture medium in the cell culture vessel to the outside in a predetermined cycle.
[0150] In the above-described first embodiment and modifications thereof, although a form in which the siphon mechanism 6a is disposed outside the temporary holding means 4 has been illustrated, for example, as shown in
[0151] Accordingly, when the height of the culture medium in the temporary holding means 4 reaches the height of the turn-around point 6b or higher, the culture medium in the temporary holding means 4 starts to be discharged from the discharge port 4b via the flow passage of the siphon mechanism 6a according to the siphon principle and keeps being discharged until the height of the culture medium in the temporary holding means 4 reaches the opening 6d of the siphon mechanism 6a. Then, the height of the culture medium in the temporary holding means 4 starts to rise due to the culture medium supplied from the culture-medium storage means 3, and, when the height of the culture medium in the temporary holding means 4 reaches a position higher than the turn-around point 6b of the flow passage of the siphon mechanism 6a, discharge occurs again. This process is repeated.
[0152] Specifically, the siphon mechanism 6a of the present invention is a mechanism that is provided with a flow passage capable of intermittently supplying the culture medium from the temporary holding means 4 to the cell culture vessel according to the siphon principle.
[0153] According to the present invention, it is possible to provide a cell culture vessel including: a supply port for supplying a culture medium; a discharge port for discharging the culture medium; and a tubular member (discharge siphon mechanism) that is connected to the discharge port, wherein the tubular member forms a flow passage that allows the culture medium in the cell culture vessel to be intermittently discharged according to the siphon principle.
[0154] For example, it is possible to provide a cell culture vessel 7, such as those shown in
[0155] Furthermore, it is possible to provide a cell culture vessel 7 such as those shown in
[0156] The culture-medium supply means 6, which supplies the culture medium to such cell culture vessels 7, is connected to the supply ports of the cell culture vessels 7, thereby making it possible to configure a cell culture system. As the culture-medium supply means 6, for example, as shown in
[0157] Furthermore, the supply port may be disposed in the top surface of the cell culture vessel 7, and the culture medium may be dripped from the supply port onto the top surface of the culture medium by the culture-medium supply means 6. By doing so, backflow of the culture medium can be prevented, thereby making it possible to prevent the culture medium in the culture-medium storage vessel, which is disposed at an upstream side of the system, from being contaminated (the occurrence of contamination).
[0158] According to this cell culture system, the culture medium is supplied to the cell culture vessels 7 by the culture-medium supply means 6, and, when the culture medium in the cell culture vessels 7 exceeds a predetermined amount, the discharge siphon mechanism 6a discharges the culture medium to a predetermined height thereof on the basis of the siphon principle, thereby making it possible to intermittently replace the old culture medium with a new culture medium.
[0159] In the above-described embodiments and the modifications thereof, a form in which the culture medium is intermittently supplied and discharged according to the siphon principle has been illustrated; however, in a cell culture system using a porous membrane, such as a dialysis membrane, a solution, such as a dialysate solution, may be intermittently supplied and discharged according to the siphon principle.
[0160]
[0161] Furthermore,
[0162] This form can be applied to floating cells as well as adherent cells.
[0163] Specifically, according to the present invention, it is possible to provide a cell culture system including:
[0164] a solution storage means that holds a solution;
[0165] a temporary holding means that is coupled to the solution storage means and that is provided with a holding space for temporarily holding the solution supplied from the solution storage means and a discharge port for discharging the solution;
[0166] a solution supply means that is coupled to the discharge port of the temporary holding means and that supplies the solution discharged from the temporary holding means, to a cell culture vessel; and
[0167] a discharge means that discharges the solution from the cell culture vessel;
[0168] wherein the cell culture vessel is coupled to the solution supply means at a position lower in the direction of gravitational force than the solution supply means; and
[0169] the solution supply means is provided with a flow passage that allows the solution to be intermittently supplied from the temporary holding means to the cell culture vessel according to the siphon principle.
[0170] Furthermore, according to the present invention, it is possible to provide a cell culture system including:
[0171] a temporary holding means that is provided with a solution storage means that holds a solution, a holding space that is coupled to the solution storage means and that temporarily holds the solution supplied from the solution storage means, and a discharge port for discharging the solution;
[0172] a solution supply means that is coupled to the discharge port of the temporary holding means and that supplies the solution discharged from the temporary holding means, to a cell culture vessel; and
[0173] a discharge means that discharges the solution from the cell culture vessel,
[0174] wherein the cell culture vessel is coupled to the solution supply means at a position lower in the direction of gravitational force than the solution supply means; and the solution supply means is provided with a supply gate that opens a flow passage when the culture medium in the holding space reaches a predetermined amount.
[0175] As a result, the above-described embodiments lead the following aspects.
[0176] According to one aspect, the present invention provides a cell culture system including: a culture-medium storage means that holds a culture medium for culturing cells; a temporary holding means that is coupled to the culture-medium storage means and that is provided with a holding space for temporarily holding the culture medium supplied from the culture-medium storage means and a discharge port for discharging the culture medium; a culture-medium supply means that is coupled to the discharge port of the temporary holding means and that supplies the culture medium discharged from the temporary holding means, to a cell culture vessel; and a discharge means that discharges the culture medium from the cell culture vessel, wherein the cell culture vessel is coupled to the culture-medium supply means at a position lower in the direction of gravitational force than the culture-medium supply means; and the culture-medium supply means is provided with a flow passage that allows the culture medium to be intermittently supplied from the temporary holding means to the cell culture vessel according to the siphon principle.
[0177] According to this aspect, even when the user is not present, a solution, such as a culture medium, can be automatically supplied to and discharged from the cell culture vessel. The siphon principle is used, thus allowing the solution, such as a culture medium, to be intermittently supplied to the cell culture vessel, with a simple configuration.
[0178] According to another aspect, the present invention provides a cell culture system including: a culture-medium storage means that holds a culture medium for culturing cells; a temporary holding means that is coupled to the culture-medium storage means and that is provided with a holding space for temporarily holding the culture medium supplied from the culture-medium storage means and a discharge port for discharging the culture medium; a culture-medium supply means that is coupled to the discharge port of the temporary holding means and that supplies the culture medium discharged from the temporary holding means, to a cell culture vessel; and a discharge means that discharges the culture medium from the cell culture vessel, wherein the cell culture vessel is coupled to the culture-medium supply means at a position lower in the direction of gravitational force than the culture-medium supply means; and the culture-medium supply means is provided with a supply gate that opens a flow passage when the culture medium in the holding space reaches a predetermined amount.
[0179] According to this aspect, even when the user is not present, a solution, such as a culture medium, can be automatically supplied to and discharged from the cell culture vessel. The gate, which opens and closes according to the amount of the culture medium in the temporary holding means, is adopted, thus allowing the solution, such as a culture medium, to be intermittently supplied to the cell culture vessel, with a simple configuration.
[0180] Furthermore, in the above-described aspect, the culture-medium supply means may be provided with the flow passage that causes the culture medium before being supplied to the cell culture vessel to pass through a position that is hi.sub.gher in the direction of gravitational force than the discharge port of the temporary holding means and that is lower in the direction of gravitational force than the maximum height of the holding space.
[0181] Furthermore, in the above-described aspect, the discharge port may be disposed in a side surface of the temporary holding means at the position of a predetermined height from a bottom surface of the temporary holding means; and the culture-medium supply means may form the flow passage from the discharge port to the outside of the temporary holding means.
[0182] Furthermore, in the above-described aspect, the culture-medium supply means may form the flow passage from the discharge port of the temporary holding means to the inside of the temporary holding means; and an end of the culture-medium supply means that is opposite from an end thereof that is connected to the discharge port of the temporary holding means may have an opening in the vicinity of the bottom surface of the temporary holding means with a gap therebetween.
[0183] Furthermore, in the above-described aspect, the supply gate may open the flow passage when the culture medium in the holding space reaches a predetermined weight or when the top surface of the culture medium in the holding space reaches a predetermined height. Accordingly, with a simple configuration, the solution, such as a culture medium, can be intermittently supplied to the cell culture vessel.
[0184] Furthermore, in the above-described aspect, the cell culture vessel may be provided with a discharge port for discharging the culture medium to the outside when the height of the top surface of the culture medium therein reaches a predetermined height. Accordingly, with a simple configuration, the solution, such as a culture medium, can be automatically supply to and discharged from the cell culture vessel.
[0185] Furthermore, in the above-described aspect, it is possible to further include a supply-rate adjusting means that controls the supply rate of the culture medium supplied from the culture-medium storage means to the temporary holding means. Accordingly, the cycle of intermittent supply of the culture medium to the cell culture vessel can be arbitrarily set.
[0186] According to still another aspect, the present invention provides a cell culture vessel including: a supply port for supplying a culture medium; a discharge port for discharging the culture medium; and a tubular member that is connected to the discharge port, wherein the tubular member forms a flow passage that allows the culture medium in the cell culture vessel to be intermittently discharged according to the siphon principle.
[0187] According to this aspect, the tubular member may form a flow passage that passes through a position that is higher in the direction of gravitational force than the discharge port and that is lower in the direction of gravitational force than the maximum height of a space in the cell culture vessel.
[0188] Furthermore, in the above-described aspect, the discharge port may be disposed in a side surface of the cell culture vessel at the position of a predetermined height from the bottom surface of the cell culture vessel; and the tubular member may form a flow passage from the discharge port to the outside of the cell culture vessel.
[0189] Furthermore, in the above-described aspect, the tubular member may form a flow passage from the discharge port to the inside of the cell culture vessel; and an end of the tubular member that is opposite from an end thereof that is connected to the discharge port may have an opening in the vicinity of the bottom surface of the cell culture vessel with a gap therebetween.
[0190] According to still another aspect, the present invention provides a cell culture system including: the above-described cell culture vessel; and a culture-medium supply means that supplies the culture medium from the supply port of the cell culture vessel to the inside thereof.
[0191] According to the present invention, it is possible to replace a culture medium even when a worker is not present and to reduce the number of times the worker walks into a work space. Accordingly, it is possible to save the trouble of having to change into disposable work clothes and the costs thereof and to reduce the risk of contamination of a cell culture system by bacteria etc. Furthermore, because the configuration of the system is simple, it is possible to reduce the risk of a system error and to avoid the risk of an impact on culture conditions due to an error.
Reference Signs List
[0192] 1 incubator
[0193] 2 temperature control means
[0194] 3 culture-medium storage means
[0195] 4 temporary holding means
[0196] 4c backup discharge port
[0197] 5 supply-rate adjusting means
[0198] 5a liquid feeding pump
[0199] 5b supply-rate adjusting means
[0200] 6 culture-medium supply means
[0201] 6a siphon mechanism
[0202] 6c branching means
[0203] 6d supply gate
[0204] 7 cell culture vessel
[0205] 8 culture-medium discharge means
[0206] 8a waste-liquid holding means
[0207] 19 control unit
[0208] 20 tubular member (tube)
[0209] 21 plate-like member
[0210] 22 spherical (columnar) member
[0211] 23, 25, 26 through-hole
[0212] 24 shutter member
[0213] 31 valve
[0214] 32 ring member
[0215] 33 ball
[0216] 34 elastic member
[0217] 41 valve
[0218] 42 float
[0219] 43 ring member
[0220] 44 ball
[0221] 45 string-like member
[0222] 51 pressure applying means
[0223] 61 saclike member
[0224] 62, 64 isolation member
[0225] 65 weight
[0226] 71 negative-pressure supply means
[0227] 81 pump
[0228] 82 waste-liquid container
[0229] 83 suction port
[0230] 91 culture vessel
[0231] 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200 cell culture system