Cell observation system and cell observation method

Abstract

To provide a cell observation system capable of obtaining an observation image of a cell in which occurrence of halation and distortion is reduced.SOLUTION: A cell observation system for observing cells adhering to a culture vessel includes a shaking unit configured to perform a shaking operation of reciprocating the culture vessel, and an observation unit configured to observe the cells when the culture vessel is moved, wherein the reciprocating movement is a movement of repeating a forward path and a return path in a trajectory connecting two points, an average speed of the culture vessel in the forward path is higher than an average speed of the culture vessel in the return path, and the observation unit observes the cells when the culture vessel is located near an end point of the return path.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to a cell observation system and a cell observation method. [Background technology]

[0002] In the medical field, cells that can be used for cell therapy and cell transplantation are cultivated. In particular, in the cultivation of adherent cells, the cells are detached from the culture vessel and seeded in another culture vessel to proliferate. Cell detachment can be achieved by using enzymes or chemicals that act on the membrane proteins that contribute to cell adhesion.

[0003] On the other hand, methods of detaching cells to improve their adhesion and survival rate after detachment, such as shaking the culture vessel or using surface-treated culture vessels, have been investigated. When detaching cells from the culture vessel using these methods, it is necessary to observe the state of the cells during the detachment process to prevent excessive stimulation of the cells.

[0004] Patent Document 1 discloses a cell detachment device that detaches cells adhered to a culture vessel by reciprocating a vessel holder with the culture vessel attached and causing it to collide with a collision target. Patent Document 2 also discloses a device that acquires images of cells cultured on a tray and measures the cell density to adjust the operation of the tray. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] Japanese Patent Application Laid-Open No. 2014-113133 [Patent Document 2] Special Publication No. 2019-526288 Summary of the Invention [Problem to be solved by the invention]

[0006] When cells are detached from a culture vessel, a solution such as a culture medium or a treatment solution for protecting the cells is generally retained in the culture vessel. During the shaking process of shaking the culture vessel, when observing the cells, the shaking of the liquid surface can cause blown-out highlights and distortions in the observed image of the cells, making it impossible to observe the cells with high resolution.

[0007] Therefore, an object of the present invention is to provide a cell observation system that can obtain an observation image of a cell with reduced occurrence of overexposure and distortion. Another object of the present invention is to provide a cell observation method that can obtain an observation image of a cell with reduced occurrence of overexposure and distortion. [Means for solving the problem]

[0008] The above object is achieved by the present invention, which provides a cell observation system for observing cells adhered to a culture vessel, comprising: a shaking means for shaking the culture vessel to move back and forth; and an observation means for observing the cells as the culture vessel is moved; the back and forth movement is a repeated movement of an outward and return path on a trajectory connecting two points; the average speed of the culture vessel on the outward path is greater than the average speed on the return path; and the observation means observes the cells when the culture vessel is located near the end point of the return path.

[0009] Furthermore, according to the present invention, there is provided a cell observation method for observing cells adhered to a culture vessel, comprising: a shaking step of performing a shaking operation to move the culture vessel back and forth; and an observation step of observing the cells during the shaking step, wherein the back and forth movement is a repeated forward and backward movement on a trajectory connecting two points, the average speed of the culture vessel on the forward path is greater than the average speed on the backward path, and the observation step includes a step of observing the cells when the culture vessel is located near the end point of the backward path. [Effects of the Invention]

[0010] According to the present invention, it is possible to provide a cell observation system that can obtain an observation image of a cell with reduced occurrence of overexposure and distortion, and a cell observation method that can obtain an observation image of a cell with reduced occurrence of overexposure and distortion. [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 is a schematic diagram showing a cell observation system according to a first embodiment. [Figure 2] 1 is a block diagram showing the configuration of a cell observation system according to a first embodiment. [Figure 3] FIG. 2 is a schematic diagram illustrating the operation of the cell observation system according to the first embodiment. [Figure 4] 3 is a schematic diagram showing the state of the liquid surface in the cell observation system according to the first embodiment. FIG. [Figure 5] FIG. 10 is a schematic diagram showing a cell observation system according to a second embodiment. [Figure 6] FIG. 10 is a schematic diagram illustrating the operation of the cell observation system according to the second embodiment. [Figure 7] FIG. 1 is a schematic diagram showing a shaking process in the prior art. [Figure 8] FIG. 2 is a diagram showing an observation image in the cell observation system according to the first embodiment. [Figure 9] 4 is a flowchart showing the operation of the cell observation system according to the first embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0012] The present invention will be described in further detail below with reference to preferred embodiments. The present invention is not limited to the following embodiments as long as the gist of the invention is not exceeded.

[0013] The present inventors have investigated methods for observing the state of cells during a shaking process in which a culture vessel to which cells have adhered is shaken, and have found that the shaking action can cause the liquid surface of the culture medium, treatment solution, etc. held in the culture vessel to shake, resulting in overexposure and distortion in the observed image.

[0014] Figure 7(a) is a schematic diagram showing a conventional shaking process. Figure 7(b) is a graph showing the amount of movement over time in a conventional shaking process. As shown in Figure 7(a), when the culture vessel moves due to the shaking operation, the liquid surface held in the culture vessel wobbles. This can cause the illumination light reflected by the liquid surface to appear in the observed image, resulting in overexposure, or can cause distortion in the observed cell image due to the influence of liquid surfaces with different heights and curvatures.

[0015] Based on the above findings, the present inventors investigated a shaking operation in which the average speed of the outward and return paths was changed and the culture vessel was moved back and forth along a fixed trajectory. As a result, the present inventors found that when the culture vessel moves from the return path, which has a slow average speed, to the outward path, which has a fast average speed, by performing observation near the end point of the return path, it is possible to achieve both ease of cell detachment and reduction in the occurrence of overexposure and distortion in the observed image.

[0016] The reason why blown-out highlights and distortions in the observed image are reduced is explained as follows. Note that, in the reciprocating motion, the movement with a high average speed will be described as the "forward path" and the movement with a low average speed as the "return path," but these movements may be repeated alternately. Either the forward path or the return path may be started first.

[0017] When a culture vessel is moved back and forth, the action of inertia causes waves on the liquid surface to become high the moment the movement in one direction is completed. If the outward movement of the reciprocating movement is assumed to have a high average speed and the return movement to have a low average speed, the waves generated on the outward movement will become smaller with the return movement. Therefore, by capturing an image of the cells near the end point of the return movement, the effects of the swaying of the liquid surface can be reduced, reducing the occurrence of overexposure and distortion.

[0018] The cell observation system and cell observation method according to the present invention will be described below with reference to specific device configurations. Note that the cell observation system and cell observation method according to the present invention are not limited to the following configurations and methods, and may be implemented in combination with other cell processing devices and methods.

[0019] [Embodiment 1] The cell observation system according to this embodiment includes a shaking means for shaking the culture vessel to move it back and forth, and an observation means for observing cells adhered to the shaking culture vessel. In this embodiment, an example will be described in which the culture vessel is moved back and forth along a horizontal, linear trajectory.

[0020] The reciprocating movement is not limited to a horizontal straight line, and any reciprocating movement that repeats forward and backward movements on a trajectory connecting two points may be used as long as it generates temporary acceleration in the culture vessel. For example, the trajectory of the reciprocating movement may be a straight line including the horizontal direction, an ellipse whose major axis is oriented in the horizontal direction, or a circular arc. The trajectory of the reciprocating movement may be selected depending on the cell type, shape, culture conditions, etc. From the perspective of increasing cell detachment efficiency, a straight trajectory including the horizontal direction is more preferable, and a horizontal straight trajectory is even more preferable, since a culture vessel placed horizontally is likely to generate large acceleration.

[0021] Furthermore, the cell observation method according to this embodiment includes a shaking step of performing a shaking operation to move a culture vessel to which cells have adhered back and forth, and an observation step of observing the cells during the shaking step. The back-and-forth movement is a movement that repeats an outward and return path on a trajectory connecting two points. The average speed of the culture vessel on the outward path is greater than the average speed on the return path. The observation step is characterized by including a step of observing the cells when the culture vessel is located near the end point of the return path.

[0022] (Outline of the cell observation system) FIG. 1 shows a schematic diagram of a cell observation system according to this embodiment. The cell observation system according to this embodiment has an agitation unit 4, which is an agitation means, a mounting unit 5 on which a culture vessel is placed, and an imaging unit 10, which is an observation means. The cell observation system according to this embodiment may be configured to include a cell detachment device 1 having the agitation unit 4 and the mounting unit 5. The agitation unit 4 is placed on a base plate 3 to which rubber feet 2 are attached. The mounting unit 5 is located above the agitation unit 4. When using the cell observation system according to this embodiment, a culture vessel 6 holding cells is placed on the mounting unit 5.

[0023] The shaking unit 4 has a driving unit 41 for shaking the mounting unit 5 and the culture vessel 6, and a detecting unit 42 for detecting the position of the mounting unit 5 during the shaking operation. The driving unit 41 may be configured to be capable of reciprocating along a linear track using, for example, a linear guide mechanism (not shown) provided on the base plate 3. Alternatively, the driving unit 41 may be configured to transmit the drive of a pulse motor (not shown) to the mounting unit 5 via a timing belt (not shown).

[0024] Above the mounting unit 5, there are arranged an imaging unit 10, which is a means for observing the cells held in the culture vessel 6, and an illumination unit 9, which illuminates the cell sheet 8 during imaging. In this embodiment, an example will be described in which the cells held in the culture vessel are observed from above in the direction of gravity of the culture vessel. By observing the cells from above, there is no need to provide an observation unit below the culture vessel, and this makes it easier to design an apparatus for performing operations such as heating and shaking.

[0025] The imaging unit 10, which is an observation means, is preferably placed in a position where the culture vessel 6 comes into view when the culture vessel 6 is at the end point of the return path. By placing the imaging unit 10 in a position suitable for imaging in advance, the cell observation system including the imaging unit 10 can be made smaller.

[0026] The culture vessel 6 includes, for example, a bottom 61 and a lid 62. The culture vessel 6 may be any vessel that allows observation of the interior from above, that is, any vessel whose lid 62 or vessel wall transmits visible light. While FIG. 1 shows the culture vessel 6 as a lidded dish as an example, other than a dish, a flask, a tray, a well plate, etc. may also be used. The culture vessel 6 may be made of, for example, polystyrene.

[0027] The culture vessel 6 is shaken while holding the cell sheet 8 and treatment solution 7 inside. Here, an example is described in which the cell sheet 8 is detached from the culture surface (bottom surface) of the culture vessel 6, but the cell observation system of the present invention can be used for purposes other than observing cell sheets. In addition to cell sheets, the culture vessel 6 may hold any of single cells, spheroids, and organoids.

[0028] The treatment liquid 7 is preferably, for example, a medium used for cell culture or a buffer solution with adjusted osmotic pressure. Among these, a medium containing amino acids, organic acids, vitamins, nucleic acids, etc. is particularly preferred from the viewpoint of increasing the survival rate of the cells that constitute the cell sheet 8.

[0029] The cell sheet 8 can be formed by a well-known culture method. A cell suspension is seeded in the culture vessel 6 at a predetermined density, and the cells are cultured in an incubator, thereby forming the cell sheet 8.

[0030] The cell observation system disclosed in this specification can be used in a cell detachment system that detaches cells from a culture vessel, and in a cell manufacturing system that detaches and recovers cells from a culture vessel.

[0031] (Operation of each part of the cell observation system) The shaking unit 4 of this embodiment performs a shaking action to move the culture vessel 6 to and fro with the cell sheet 8 adhered thereto, thereby detaching the cell sheet 8 from the culture vessel 6. The photographing unit 10, which is an observation means, observes the cell sheet 8 during the shaking action.

[0032] FIG. 2 shows a block diagram of the cell observation system of this embodiment. The shaking unit 4 includes a drive unit 41 that moves the mounting unit 5 back and forth in the X direction, and a detection unit 42 that detects the position of the mounting unit 5. For example, when the drive unit 41 uses a pulse motor as a drive source to shake the culture vessel 6 via the mounting unit 5 using a timing belt and a linear guide (not shown), the pulse motor is driven forward and backward to achieve the desired back and forth movement. Furthermore, a position sensor, which is an example of the detection unit 42, detects the position of the mounting unit 5 on the linear guide. The detection unit 42 may also calculate the back and forth period of the shaking operation.

[0033] The control unit 11 instructs the photographing unit 10 to photograph in accordance with the position of the mounting unit 5 detected by the detection unit 42. At this time, the control unit 11 may instruct the photographing unit 10 to photograph immediately, or may instruct the photographing unit 10 to photograph after a certain time has elapsed so that photographing can be performed at a preferred position.

[0034] Furthermore, the image of the cell sheet 8 captured by the imaging unit 10 may be used by the data processing unit 12 to determine whether or not the cell sheet 8 has detached. The determination can be made, for example, by threshold determination based on the brightness distribution of the image, or by determination based on information about an image for determination stored in advance in a storage means (not shown). From the viewpoint of reducing tearing of the cell sheet 8, it is more preferable to control the shaking unit 4 to stop driving based on the determination of detachment of the cell sheet 8.

[0035] (Cell observation method) Figure 3 shows a schematic diagram illustrating the operation of the cell observation system according to this embodiment. Figure 3(a) shows the movement of the culture vessel 6 as seen from the side. Figure 3(b) shows the relationship between the movement amount (amplitude) in the X direction and the time axis during the shaking operation.

[0036] (i) in Figure 3(a) shows a state in which the culture vessel 6 is located at the start point of the forward path. Then, in (ii), the culture vessel 6 is moved along the forward path at an average speed V1. Movement at the average speed V1 promotes detachment of the cell sheet 8 held in the culture vessel 6. In (iii), the culture vessel 6 reaches the end point of the forward path. In (iv), the culture vessel 6 is moved along the return path at an average speed V2 that is slower than the average speed V2. The cell detachment method according to this embodiment involves repeatedly moving the culture vessel 6 along the forward and return paths shown in (i) to (iv) as a shaking operation.

[0037] The average speed here is the value (mm / sec) obtained by dividing the movement amount (amplitude movement amount 2T [mm]) of the culture vessel 6 between the start point and end point of either the forward or return path of the shaking motion trajectory by the time required for the movement. The average speed V1 of the forward path should be greater than the average speed V2 of the return path, and the movement of the culture vessel 6 may be decelerated near either the start point or end point of the forward or return path.

[0038] In cell sheet detachment, different average speeds for the forward and backward movements can also be used to reduce tearing and damage to the cell sheet. When the cell sheet is repeatedly shaken after its edge is turned up by the shaking action, the treatment solution penetrates between the edge and the culture vessel, promoting detachment. On the other hand, the flow of treatment solution may collide with the cell sheet from the opposite direction to the turned-up edge, causing tearing and damage to the cell sheet. Therefore, by performing the forward movement, which rapidly promotes detachment, and the backward movement, which reduces damage, at different average speeds, efficient detachment can be achieved while reducing tearing and damage to the cell sheet.

[0039] Furthermore, when peeling a cell sheet, it is more preferable to change the direction of reciprocating movement depending on the state of the cell sheet acquired by the observation means. For example, referring to FIG. 3(a), if the cell sheet on the -X side is beginning to peel, it is more preferable to perform reciprocating movement so that a fast flow of treatment solution is directed toward the end of the cell sheet where peeling has begun. That is, it is more preferable to move the culture vessel in the direction of peeling beginning (-X direction) as the outward movement with a higher average speed, and then move the culture vessel in the +X direction as the return movement with a slower average speed than the outward movement. By processing the observed image of the cell sheet using well-known techniques, it is possible to analyze the direction from which the cell sheet is beginning to peel. By performing a shaking operation in accordance with the acquired peeling direction, the treatment solution enters between the turned-up cell sheet and the culture vessel, thereby efficiently peeling the cell sheet while reducing tearing and damage.

[0040] FIG. 3(b) shows the relationship between the movement amount and the time axis of the shaking operation in this embodiment. Here, the distance from the midpoint of the forward and backward movements is shown as the movement amount (amplitude). When the forward movement starts, waves of the treatment solution 7 held in the culture vessel 6 are generated with a delay (15d in FIG. 3(a)), and the waves of the treatment solution 7 become larger near the midpoint (ii) of the forward movement (15a in FIG. 3(a)). When the culture vessel 6 reaches the end point (iii) of the forward movement and starts the backward movement, the waves of the treatment solution 7 collide with the vessel wall of the culture vessel 6 (15b in FIG. 3(a)).

[0041] Next, the culture vessel 6 is moved on the return path at an average speed V2. Since the average speed of the return path is slower than that of the forward path, the waves of the treatment solution 7 generated during the forward path are smaller (15c in FIG. 3(a)). In the shaking operation of this embodiment, the above-mentioned forward and return paths are repeated.

[0042] In the cell observation method of this embodiment, during the shaking operation described above, cells held in the culture vessel 6 are observed when the culture vessel 6 is positioned near the end point of the return path. By positioning the culture vessel 6 near the end point of the return path, the cells can be observed with small waves in the treatment solution 7, reducing overexposure in the observed image. It is more preferable to observe the cells when the culture vessel 6 is positioned between the midpoint (iv) of the return path and the midpoint (ii) of the forward path. In particular, it is more preferable to observe the cells when the culture vessel 6 is positioned between the midpoint (iv) of the return path and the midpoint between the start point (i) and the midpoint (ii) of the forward path.

[0043] Observation of the cell sheet 8 by the photographing unit 10, which is the observation means, is performed with the culture vessel 6 positioned near the end point of the return path, as described above. From the perspective of efficiently observing the cell sheet 8, it is preferable that observation by the photographing unit 10 be performed in a time shorter than the time it takes for the culture vessel 6 to make one round trip. In particular, it is preferable that observation be performed in a time shorter than half the time it takes for the culture vessel 6 to make one round trip between the forward and return paths. Note that observation may be performed by photographing at a desired shutter speed, or by extracting an image at a desired time from video-captured data.

[0044] By performing the observation near the end point of the return path, the image quality of the observation image can be improved. Near the end point of the return path, the moving speed of the culture vessel 6 is relatively slow, so image blur due to motion blur can be reduced. In addition, the light intensity of the observation light source can be reduced, so whiteout due to reflection on the liquid surface can be reduced.

[0045] 9 is a flowchart showing the cell observation method according to this embodiment. In S101, the shaking operation by the shaking unit 4 is started. In S102, the control unit 11 determines whether to perform observation based on information acquired by the detection unit 42. In S103, the control unit 11 instructs the imaging unit 10 to perform observation.

[0046] (Example) The results of an experiment using the cell observation system will be explained using Figures 4 and 8. A 35 mm diameter dish was used as the culture vessel. 2 cc of medium was poured into the dish and cells were seeded. The culture vessel was incubated in an incubator at 37°C for several days to culture the cells and form a cell sheet. The culture vessel was removed from the incubator and placed on the mounting section of the cell observation system of this embodiment, and a shaking operation was performed. The amplitude of the shaking operation was 25 mm, the outgoing frequency was 1.66 Hz, and the return frequency was 1.25 Hz.

[0047] Figures 4 and 8 show images of waves in the culture medium in a culture vessel during shaking. On the outbound path, the average speed was high, so inertial forces were generated in the culture medium due to sudden acceleration, resulting in large waves. Furthermore, at the end of the outbound path, the culture medium, acting on the inertial forces, collided with the inner wall of the culture vessel, causing the waves to become even larger. At the midpoint of the return path, the waves that had occurred on the outbound path became smaller. Because the average speed on the return path was low, the waves at the end of the return path were small. From these results, it was found that the waves were small between the midpoint of the return path and the midpoint of the outbound path, making it ideal for observation. In particular, it was found that the waves were particularly small between the midpoint of the return path and the midpoint between the start and midpoint of the outbound path.

[0048] As a result of the cell sheet detachment experiment using shaking, the cell sheet was completely detached within about 30 minutes of starting the shaking.

[0049] [Embodiment 2] The cell observation system according to this embodiment performs a shaking motion along an arcuate trajectory that includes horizontal movement, and observes the cells while detaching them from the flask container. The cell observation system according to this embodiment will be described using Figures 5 and 6. Explanations of parts common to embodiment 1 will be omitted.

[0050] The cell observation system of this embodiment performs a shaking operation by moving the mounting part 5 back and forth along an arc orbit. The mounting part 5 is moved by driving the shaking motor 22. By controlling the driving of the shaking motor 22, the shaking speed (frequency) of the outward and return paths can be adjusted.

[0051] In this embodiment, an example is shown in which a flask 20 is used as the culture vessel. The flask 20 is composed of a body 20a and a screw-type lid 20b. By closing the lid of the flask 20, leakage of the treatment solution during the shaking operation can be prevented, and the conditions for the shaking operation can be set widely.

[0052] The illumination unit 9 and the photographing unit 10 are disposed, for example, between the rotation fulcrum 21 and the mounting unit 5. The illumination unit 9 and the photographing unit 10 are preferably disposed at an angle offset from the center of the shaking operation in advance so that the culture vessel in a position suitable for photographing comes into view.

[0053] 6 is a diagram showing the shaking operation according to this embodiment. From the state in which flask 20 moves in the +X direction in (i), flask 20 moves on the outward path at rotational speed W1 and reaches position (iii). At this time, waves are generated within flask 20 due to the movement at rotational speed W1, but the waves become smaller as flask 20 moves on the return path at rotational speed W2, which is slower than rotational speed W1. Note that the rotational speed here refers to the average speed on either the outward or return path.

[0054] In this way, by differentiating the rotation speeds of the forward and backward movements, it is possible to efficiently detach the cell sheet while reducing tearing and damage for the same reasons as described above. Furthermore, using a flask can prevent leakage of the treatment solution (culture medium).

[0055] The disclosure of this embodiment includes the following configurations and methods.

[0056] (Configuration 1) A cell observation system for observing cells adhered to a culture vessel, comprising: a shaking means for performing a shaking operation by moving the culture vessel back and forth; an observation means for observing the cells when the culture vessel is moved; The reciprocating movement is a movement that repeats a forward path and a backward path on a track connecting two points, an average speed of the culture vessel on the outward path is greater than an average speed of the culture vessel on the return path; The cell observation system is characterized in that the observation means observes the cells when the culture vessel is located near the end point of the return path.

[0057] (Configuration 2) The cell observation system described in Configuration 1, characterized in that the time during which the observation means observes the cells while the culture vessel makes one round trip between the outbound path and the return path is shorter than half the time it takes for the culture vessel to make one round trip between the outbound path and the return path.

[0058] (Configuration 3) 3. The cell observation system according to configuration 1 or 2, wherein the observation means observes the cells from above in the direction of gravity of the culture vessel.

[0059] (Configuration 4) 4. The cell observation system according to any one of configurations 1 to 3, wherein the trajectory is a straight line extending in a direction including the horizontal direction.

[0060] (Configuration 5) 5. The cell observation system according to configuration 4, wherein the trajectory is a horizontal straight line.

[0061] (Configuration 6) 4. The cell observation system according to any one of configurations 1 to 3, wherein the orbit is an ellipse whose major axis includes the horizontal direction.

[0062] (Configuration 7) 4. The cell observation system according to any one of configurations 1 to 3, wherein the orbit is an arc.

[0063] (Configuration 8) 8. The cell observation system according to any one of configurations 1 to 7, wherein the observation means is disposed at a position where the culture vessel located at the end point of the return path is within a field of view.

[0064] (Configuration 9) The cells are a cell sheet, 2. The cell observation system according to configuration 1, wherein the shaking means changes the direction of the reciprocating movement depending on the state of the cell sheet acquired by the observation means.

[0065] (Configuration 10) 10. A cell detachment system comprising the cell observation system according to any one of configurations 1 to 9, wherein the cells are detached from the culture vessel by the shaking means.

[0066] (Configuration 11) A cell manufacturing system comprising the cell observation system according to any one of configurations 1 to 9, wherein the cells are detached from the culture vessel by the shaking means and the cells are recovered.

[0067] (Method 1) A cell observation method for observing cells adhered to a culture vessel, comprising: a shaking step of performing a shaking operation in which the culture vessel is moved back and forth; an observation step of observing the cells during the shaking step, The reciprocating movement is a movement that repeats a forward path and a backward path on a track connecting two points, an average speed of the culture vessel on the outward path is greater than an average speed of the culture vessel on the return path; A cell observation method, characterized in that the observation step includes a step of observing the cells when the culture vessel is located near the end point of the return path.

[0068] (Method 2) A method for producing cells, comprising detaching the cells from the culture vessel using the cell observation method described in Method 1, and recovering the cells. [Explanation of symbols]

[0069] 4. Shaking section 5. Placement section 6 Culture vessel 7. Culture medium 8 Cell Sheet 10. Photography Department

Claims

1. A cell observation system for observing cells adhered to a culture vessel, comprising: a shaking means for performing a shaking operation by moving the culture vessel back and forth; an observation means for observing the cells when the culture vessel is moved; The reciprocating movement is a movement that repeats a forward path and a backward path on a track connecting two points, an average speed of the culture vessel on the outward path is greater than an average speed of the culture vessel on the return path; The cell observation system is characterized in that the observation means observes the cells when the culture vessel is located near the end point of the return path.

2. The cell observation system according to claim 1, characterized in that the time during which the observation means observes the cells while the culture vessel makes one round trip between the outgoing path and the returning path is shorter than half the time during which the culture vessel makes one round trip between the outgoing path and the returning path.

3. 2. The cell observation system according to claim 1, wherein the observation means observes the cells from above the culture vessel in the direction of gravity.

4. The cell observation system according to claim 1 , wherein the trajectory is a straight line extending in a direction including the horizontal direction.

5. 5. The cell observation system according to claim 4, wherein the trajectory is a horizontal straight line.

6. The cell observation system according to claim 1 , wherein the orbit is an ellipse whose major axis includes the horizontal direction.

7. The cell observation system according to claim 1 , wherein the trajectory is an arc.

8. 2. The cell observation system according to claim 1, wherein the observation means is disposed at a position where the culture vessel located at the end point of the return path is within a field of view.

9. The cells are a cell sheet, 2. The cell observation system according to claim 1, wherein the shaking means changes the direction of the reciprocating movement depending on the state of the cell sheet acquired by the observation means.

10. A cell detachment system comprising the cell observation system according to claim 1 , wherein the cell detachment system detaches the cells from the culture vessel by the shaking means.

11. A cell manufacturing system comprising the cell observation system according to any one of claims 1 to 9, wherein the cells are detached from the culture vessel by the shaking means and the cells are recovered.

12. A cell observation method for observing cells adhered to a culture vessel, comprising: a shaking step of performing a shaking operation in which the culture vessel is moved back and forth; an observation step of observing the cells during the shaking step, The reciprocating movement is a movement that repeats a forward path and a backward path on a track connecting two points, an average speed of the culture vessel on the outward path is greater than an average speed of the culture vessel on the return path; A cell observation method, characterized in that the observation step includes a step of observing the cells when the culture vessel is located near the end point of the return path.

13. A method for producing cells, comprising: detaching the cells from the culture vessel using the cell observation method according to claim 12; and recovering the cells.

Images