Cell treatment method, cell treatment system, and cell treatment device

Abstract

To provide a cell treatment method capable of adjusting a temperature of a member contacting a culture vessel when peeling cells adhered on a culture surface of the culture vessel within a desirable range.SOLUTION: A cell treatment method in which cells adhered on a culture surface of a culture vessel are peeled from a culture vessel by using a cell treatment system equipped with a vibration part which generates vibration includes: a heating step of generating vibration of a first mode in the vibration part in a state where the vibration part and a vibration transmission material are in contact with each other to raise a temperature of the vibration transmission material; and a peeling step of peeling the cells from the culture vessel by generating vibration of a second mode different from the first mode by the vibration part in a state where the vibration transmission material and the culture vessel are in contact with each other, after starting the heating step.SELECTED DRAWING: Figure 1

Description

[Technical field]

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

[0002] In the fields of drug discovery and regenerative medicine, cells produced by cell culture are used. In particular, when mass culturing adhesive cells, a subculture operation is performed in which cells adhered to a culture vessel or scaffold material are detached and collected, then transferred to a culture vessel and cultured again. Even if a subculture operation is not performed, when using adhesive cells, it is necessary to detach the cells from the culture vessel or scaffold material. Here, since cells have a range of temperatures at which their proliferation rate and survival rate are high depending on the cell type, the process of culturing cells is generally performed in a device such as an incubator that is controlled at a constant temperature. However, when performing an operation such as cell detachment, it is necessary to remove the culture vessel from the incubator, and there is a possibility that the survival rate of the cells will decrease as the temperature of the cells in the culture vessel placed outside the incubator approaches room temperature.

[0003] Patent Document 1 discloses a culture treatment device in which a mounting table for mounting a culture vessel removed from a culture chamber is equipped with a heater, and the medium in the culture vessel is maintained at a constant temperature. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2005-328726 A Summary of the Invention [Problem to be solved by the invention]

[0005] Here, Patent Document 1 does not disclose the type of heater or the path of heat transfer from the heater to the culture vessel. Simply placing the culture vessel on a mounting table equipped with a heater may take time for the heat from the heater to be transferred to the culture vessel, and the temperature of the cells may not be sufficiently maintained.

[0006] Therefore, an object of the present invention is to provide a cell processing method capable of adjusting the temperature of a member in contact with a culture vessel within a desired range when detaching cells adhered to the culture surface of the culture vessel. Another object of the present invention is to provide a cell processing system and a cell processing device capable of adjusting the temperature of a member in contact with a culture vessel within a desired range when detaching cells adhered to the culture surface of the culture vessel. [Means for solving the problem]

[0007] The above object is achieved by the present invention as follows. That is, according to the present invention, there is provided a cell processing method for detaching cells adhered to a culture surface of a culture vessel from the culture vessel using a cell processing system equipped with a vibration unit that generates vibrations, the cell processing method comprising: a heating step in which, with the vibration unit and a vibration transmitting material in contact with each other, the vibration unit generates a vibration of a first mode to raise the temperature of the vibration transmitting material; and a detachment step in which, after the start of the heating step, with the vibration transmitting material and the culture vessel in contact with each other, the vibration unit generates a vibration of a second mode different from the first mode to detach the cells from the culture vessel.

[0008] Furthermore, according to the present invention, there is provided a cell processing device that detaches cells adhered to a culture surface of a culture vessel from the culture vessel, the cell processing device having a vibration unit that generates vibrations and a control unit that controls the vibration of the vibration unit, wherein the control unit transmits vibrations to the vibration transmission material by vibrating the vibration unit in a first mode when the vibration transmission material is in contact with the vibration transmission material, thereby increasing the temperature of the vibration transmission material, and detaches the cells from the culture vessel by vibrating the vibration unit in a second mode different from the first mode when the vibration transmission material and the culture vessel are in contact with each other.

[0009] Furthermore, according to the present invention, there is provided a cell processing system for detaching cells adhered to a culture surface of a culture vessel from the culture vessel, the cell processing system comprising a vibration unit that generates vibrations and a control unit that controls the vibration of the vibration unit, the control unit vibrating the vibration unit in a first mode when the vibration unit is in contact with a vibration transmission material, thereby transmitting vibrations to the vibration transmission material and increasing the temperature of the vibration transmission material, and vibrating the vibration unit in a second mode different from the first mode when the vibration transmission material and the culture vessel are in contact with each other, thereby detaching the cells from the culture vessel. Effect of the Invention

[0010] According to the present invention, it is possible to provide a cell processing method capable of adjusting the temperature of a member in contact with a culture vessel within a desired range when detaching cells adhered to the culture surface of the culture vessel. Also, according to the present invention, it is possible to provide a cell processing device and a cell processing system capable of adjusting the temperature of a member in contact with a culture vessel within a desired range when detaching cells adhered to the culture surface of the culture vessel. [Brief description of the drawings]

[0011] [Figure 1] FIG. 1 is a schematic diagram of a device configuration of a cell processing system according to a first embodiment. [Diagram 2] FIG. 1 is a flow diagram of a cell treatment method according to a first embodiment. [Diagram 3] FIG. 1 is a conceptual diagram of a cell treatment method according to a first embodiment. [Figure 4] FIG. 1 is a schematic cross-sectional view of a cell processing system according to a first embodiment. [Diagram 5] FIG. 3 is a conceptual diagram of heat generation according to the first embodiment. [Figure 6] 3A to 3C are schematic diagrams of vibration modes according to the first embodiment. [Figure 7] FIG. 4 is a schematic diagram of temperature change according to the first embodiment. [Figure 8] 13A and 13B are schematic diagrams of local vibration according to the second embodiment. [Figure 9]11A to 11C are schematic diagrams of vibration modes according to a second embodiment. [Figure 10] 13A to 13C are schematic diagrams illustrating an example of switching of vibration modes according to the second embodiment. [Figure 11] FIG. 13 is a schematic diagram of a configuration example of a cell processing apparatus according to a third embodiment. [Figure 12] FIG. 11 is a schematic diagram of a device configuration of a cell processing system according to a third embodiment. [Figure 13] FIG. 11 is a flow diagram of a cell treatment method according to a third embodiment. [Figure 14] FIG. 1 is a flow diagram of a cell treatment method according to a first embodiment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] The present inventors have studied a method for maintaining the temperature of cells when the cells are detached from a culture vessel. As a result, it has been found that simply providing a general heater inside the housing of a cell processing device takes time to heat up the heater, and the temperature of the vibration transmitting material on which the culture vessel is placed may not be raised sufficiently.

[0013] Based on the above findings, further investigation was carried out, and it was found that the vibration transmitting material can be heated by applying vibration to the vibration transmitting material before placing the culture vessel. It was also found that the temperature of the vibration transmitting material can be increased more quickly by directly heating the vibration transmitting material by the vibration of the vibration transmitting material than by driving only the heater.

[0014] The present invention will be described in further detail below with reference to preferred embodiments.

[0015] [First embodiment] The cell processing method according to this embodiment transmits the vibration of a vibration unit of a cell processing system that generates vibrations to a culture vessel via a vibration transmission material, thereby detaching cells that have adhered to the culture surface of the culture vessel. Furthermore, in the cell processing method according to this embodiment, in order to adjust the temperature of the cells in the culture vessel, the vibration unit is vibrated in a mode for increasing the temperature of the vibration transmission material before the culture vessel is brought into contact with the vibration transmission material. This makes it possible to manage the temperature of the cells in the culture vessel even when the culture vessel is moved from a culture environment such as an incubator to the cell processing system.

[0016] (Cell Processing System) FIG. 1 shows a schematic diagram of the device configuration of the cell processing system according to this embodiment. The cell processing system according to this embodiment has at least a vibration unit that generates vibrations and a control unit that controls the vibration of the vibration unit. The cell processing system detaches the cells 3 from the culture surface by transmitting the vibrations generated by the vibration unit to the cells 3 attached to the culture surface of the culture vessel 100 via a vibration transmission material (not shown). The control circuit in the figure is an example of the control unit, and controls the vibration of the vibration unit. The control unit may be configured with a module executed by a CPU or MPU, or may be configured with a circuit that realizes a specific function such as an ASIC.

[0017] The present invention may be realized by a cell processing system to which a plurality of devices and computing devices are connected, and a cell processing device consisting of a single device that realizes similar functions is also included in the present invention.

[0018] (Vibration part) Fig. 4 shows a schematic cross-sectional view of the cell processing system according to this embodiment. (a) shows the state before the culture vessel comes into contact with the cell processing system. The diagram shows the case where the vibration section is of a piezoelectric type. The vibration section 107 includes a vibration plate 102 and an oscillator 117 that are bonded together. When a driving voltage is applied to the piezoelectric material included in the oscillator 117, the oscillator 117 vibrates stretching and contracting. As the oscillator 117 stretches and vibrates, the vibration plate 107 performs an expanding displacement operation, i.e., a bending deformation operation, so that the vibration section 107 vibrates and generates an elastic wave.

[0019] The vibration unit may be any device capable of generating vibrations, such as a piezoelectric device or a motor. In particular, a configuration capable of generating vibrations in the ultrasonic band is preferable in order to detach cells from the culture vessel. The transducer used in the vibration unit may be of either a piezoelectric or electromagnetic type, as long as it can generate vibrations of a constant frequency. From the viewpoint of controlling the vibration waveform and vibration frequency with high precision, it is particularly preferable that the transducer is of a piezoelectric type. If it is of a piezoelectric type, the transducer can be selected from a unimorph type, a bimorph type, and a bolt-tightened Langevin type.

[0020] (Housing) The cell processing system further has a housing 101 that holds a vibration unit 107. As shown in FIG. 4, the housing 101 preferably has a bank-like protrusion above the vibration unit 107 so that the vibration transmission material can be held when the cell processing system is placed on a laboratory table or the like. In addition, a heater for increasing the temperature of the entire housing and maintaining an equilibrium state may be provided inside the housing. Both the heater that maintains the temperature of the entire housing and the vibration unit that increases the temperature of the vibration transmission material are responsible for temperature management, so that temperature adjustment can be performed quickly and efficiently when cell processing is desired.

[0021] (Vibration transmission material) When the cell processing system is used, the vibration transmission material 129 is arranged so that the vibration transmission material 129 is in contact with the vibration unit 107. The vibration transmission material may be any material capable of transmitting at least a part of the vibration generated in the vibration unit from the vibration unit to the culture vessel. From the viewpoint of transmission performance, the vibration transmission material is preferably a liquid or solid. It is more preferable that the vibration transmission material contains water or silicone rubber. By vibrating the vibration unit while the vibration transmission material is in contact with both the vibration unit and the culture vessel, the vibration can be effectively applied to the cells.

[0022] (Culture container) The culture vessel used in this embodiment may have any configuration that can be used for cell culture. In addition, a culture vessel that contains a scaffold material such as a microcarrier or a hollow fiber material may be used. Examples of the culture vessel include a dish, a flask, a well plate, and a culture bag.

[0023] The material of the culture vessel may be any material capable of transmitting vibrations or capable of deforming in response to vibrations. For example, the culture vessel may be made of a polymer material such as polystyrene, or may be made of metal. The culture vessel is disposed so as to be in contact with a vibration transmitting material disposed on the vibration unit. Here, it is preferable from the viewpoint of efficient cell detachment that the surface where the vibration transmitting material and the culture vessel come into contact is the outer surface of the culture vessel as viewed from the culture surface where the cells are adhered in the culture vessel.

[0024] (cell) The cell treatment method according to the present embodiment can be applied to various kinds of cells. Examples of the cells include Chinese hamster ovary-derived CHO cells, mouse connective tissue L929 cells, mouse skeletal muscle myoblasts (C2C12 cells), human fetal lung-derived normal diploid fibroblasts (TIG-3 cells), human fetal kidney-derived cells (HEK293 cells), human alveolar basal epithelial adenocarcinoma-derived A549 cells, human cervical cancer-derived HeLa cells, epithelial cells, endothelial cells, skeletal muscle cells, smooth muscle cells, cardiomyocytes, neuronal cells, glial cells, fibroblasts, hepatic parenchymal cells, hepatic non-parenchymal cells, adipocytes, induced pluripotent stem (iPS) cells, embryonic stem (ES) cells, embryonic germ (EG) cells, embryonic carcinoma (EC) cells, mesenchymal stem cells, hepatic stem cells, pancreatic stem cells, skin stem cells, muscle stem cells, germ stem cells, progenitor cells of each tissue, and cells induced to differentiate therefrom.

[0025] (Vibration mode) A schematic diagram of the vibration mode according to this embodiment is shown in Fig. 6. In this embodiment, the vibration unit is vibrated in at least two modes: a first mode for increasing the temperature of the vibration transmitting material, and a second mode for detaching the cells from the culture vessel.

[0026] The vibrations in the first and second modes are preferably ultrasonic vibrations for efficient temperature control and peeling. In addition, the first mode (heating vibration) has a higher frequency than the second mode (peeling vibration), so that temperature adjustment can be performed more quickly. For example, the second mode may be vibrations with a frequency sweeping from 20 kHz to 200 kHz. By using a sweep frequency, vibrations at a resonant frequency can be generated, and vibrations with a large amplitude can be obtained. In addition, for example, the first mode may be vibrations with a fixed frequency in the range from 100 kHz to 200 kHz. By using a fixed and relatively high frequency vibration, heating can be performed more quickly.

[0027] (Flow of cell processing method) Fig. 2 shows a flow of the cell processing method according to this embodiment. First, in a preparation step, the user installs a vibration transmitting material so that the vibration transmitting material is in contact with the vibration unit of the cell processing system, and prepares the vibration unit and the vibration transmitting material. Note that the vibration transmitting material may be installed in advance, or the vibration transmitting material may be fixed to the vibration unit in advance. Preparation of the vibration unit and the vibration transmitting material may be performed automatically by any system.

[0028] Next, the user inputs a command to perform preheating to the cell processing system through the input means. In response to this command, the control unit of the cell processing system commands the vibration unit to set driving conditions and starts vibration in the first mode as a heating step. In the heating step, as shown in FIG. 2(a), the vibration unit vibrates in the first mode, vibrating the vibration transmitting material and increasing the temperature of the vibration transmitting material. FIG. 5 shows a conceptual diagram of heat generation. Elastic waves generated from the vibration unit are transmitted to the vibration transmitting material, generating intermolecular frictional heat, which increases the temperature of the vibration transmitting material.

[0029] The temperature of the vibration transmission material is preferably set to a temperature suitable for the survival of the cell type of the target cell to be detached. Although the optimal temperature varies depending on the cell type, the temperature of the vibration transmission material can be set to 37°C, for example.

[0030] The command to perform preheating does not necessarily have to be issued by the user, and may be automatically issued by the cell processing system. The input means used by the user may be, for example, a button, a touch panel, a keyboard, a mouse, a controller, etc. If the housing is configured to include a heater, the heater may be started together with the start of the vibration unit in response to a command to perform preheating, as shown in the flow diagram of FIG. 14.

[0031] Fig. 7 shows an example of temperature change during the heating process. For example, if the housing has a heater, the temperature of the vibration transmitting material rises gradually as the heater temperature rises after the heater is turned on, but it takes time to reach an equilibrium temperature. Therefore, by simultaneously carrying out a heating process using the first mode vibration, the temperature of the vibration transmitting material can be raised more quickly.

[0032] Here, the cell processing system may include a measuring unit and an alarm unit. When the heating process is completed, i.e., when the temperature of the vibration transmitting material reaches a predetermined temperature, the alarm unit (not shown) may be configured to alarm those around. The alarm unit may be configured to alarm by at least one of light, sound, and text display. The temperature of the vibration transmitting material is measured by a measuring unit (not shown). By the alarm unit notifying the completion of pre-heating, the user can easily understand the timing for performing the placement process, and can work efficiently.

[0033] Subsequently, in the positioning step performed after the start of the heating step, the culture vessel is positioned so as to contact the vibration transmitting material. This positioning step results in the vibration unit, the vibration transmitting material, and the culture vessel being positioned in this order. The positioning step may be performed either manually or automatically.

[0034] In the detachment step after the arrangement step, the control unit causes the vibration unit to vibrate in the second mode, i.e., the vibration unit generates vibration in the second mode while the vibration transmitting material and the culture vessel are in contact with each other. This vibration vibrates the cells in the culture vessel, and the cells are detached from the culture surface of the culture vessel.

[0035] [Second embodiment] In this embodiment, an example of a vibration mode in the present invention will be described.

[0036] (First mode) From the viewpoint of rapid temperature rise of the vibration transmitting material, it is preferable that the first mode for raising the temperature of the vibration transmitting material is a vibration in which the amplitude of multiple different parts on the vibration surface of the vibration part is maximized. Fig. 8 is a diagram showing local vibrations on the vibration surface (diaphragm) of the vibration part, showing a state in which there are multiple places where the amplitude is maximized. By vibrating the diaphragm in a mode in which multiple local vibrations appear, the change in the shape of the vibration transmitting material in contact with the diaphragm becomes large, and the temperature of the vibration transmitting material can be raised quickly.

[0037] (Second mode) FIG. 9 is a schematic diagram of the vibration mode.

[0038] In the peeling process, the first mode and the second mode may be performed simultaneously as shown in FIG. 9(c). In other words, the second mode in the peeling process may be a superposition of a plurality of vibration modes. The second mode may include a vibration mode with a higher frequency than the first mode, so that the temperature of the vibration transmitting material can be effectively increased. The second mode may be a square wave as shown in FIG. 9(a) or a triangular wave as shown in FIG. 9(b).

[0039] By performing vibration in the above-mentioned second mode, the temperature of the vibration transmitting material can be increased even after the culture vessel has been placed, and the temperature of the cells can be adjusted.

[0040] (Switch vibration mode) Fig. 10 is a diagram showing an example of switching of vibration modes. As shown in Fig. 10(b), by performing the heating process after the start of the peeling process, the temperature of the vibration transmitting material can be increased again in cases where the temperature of the vibration transmitting material decreases due to the vibration of the second vibration mode alone.

[0041] In addition, in the heating step, it is preferable that the temperature of the vibration transmitting material is measured by a measuring unit (not shown), and when the temperature of the vibration transmitting material is out of a predetermined range, the control unit changes the vibration frequency or amplitude of the vibration unit. By changing the vibration frequency or amplitude, it is possible to maintain the temperature of the vibration transmitting material at a temperature suitable for cell survival. For example, when it is desired to maintain the temperature of the vibration transmitting material at 37°C, it is effective to control the amplitude to be small when the measured temperature is 39°C and to be large when the measured temperature is 35°C. Note that the object for which the measuring unit measures the temperature is not limited to the vibration transmitting material, and may be a part of the vibration unit.

[0042] [Third embodiment] (Example of the configuration of a cell processing device) A schematic diagram of a configuration example of a cell processing device according to this embodiment is shown in Fig. 11. The housing 101 of the cell processing device may be provided with a power button 126, a peel button 127, and a completion button 128. By providing such buttons, the device can be used by itself, and a simple and space-saving cell processing device can be provided.

[0043] As shown in Fig. 3, the use of the cell processing device using the power button etc. is, for example, carried out in the following flow. When the power button 126 is pressed, the heater that warms the housing 101 with a constant input is turned ON, and the heating vibration of the vibration unit 107 (vibration in the first mode) begins. After that, when a preset heating time has elapsed, the heating vibration is stopped. At this time, the heater is maintained in the power ON state.

[0044] Next, the culture vessel 100 removed from the incubator is placed in the cell processing device, and the detachment vibration (second mode vibration) starts when the detachment button 127 is pressed. After that, when it is visually confirmed that the cells have been detached, the completion button 128 is pressed, and the detachment vibration stops.

[0045] When performing cell detachment from multiple culture vessels in succession, the second and subsequent culture vessels are placed in the cell processing device immediately after the first one has been processed. After placing the culture vessel, the detachment button 127 is pressed, and after confirming the detachment, the completion button 128 is pressed, and this process is repeated. When cell detachment from all the culture vessels is completed, the power button is pressed again to turn it OFF and stop the heater.

[0046] (Example of temperature control mechanism) FIG. 12 shows a schematic diagram of the device configuration of the cell processing system according to this embodiment. In order to prevent the cells from rising rapidly or becoming too hot, it is preferable to have a temperature adjustment mechanism as shown in FIG. 12. A cooling space is provided below the vibration unit 107, i.e., on the opposite side to the culture vessel. The cooling space 135 is a space covered with a wall of a heat insulating material 132 in contact with the housing 101, and partitioned by a cooling plate 130 with good thermal conductivity at the bottom. The cooling plate 130 abuts against a cooler such as a Peltier element and is provided with a cooling fin 104 on the heat exhaust side. A cooling fan 133 for blowing exhaust heat is installed under the cooling fin. By providing such a temperature adjustment mechanism, the temperature of the cells can be controlled more accurately.

[0047] (Example of a cell processing system having a memory unit) The cell processing system according to this embodiment may have a storage unit. The storage unit can be configured using any storage medium such as a memory or an optical disk. Fig. 13 shows a flow diagram of the cell processing method according to this embodiment. The storage unit stores processing conditions linked to at least one of the cell type, the cell culture conditions, and the type of culture vessel.

[0048] The processing conditions include at least one of temperature, vibration frequency, and driving voltage. The control unit controls the vibration of the vibration unit by referring to the processing conditions linked to the identification information stored in the memory unit according to the identification information (ID) of the culture vessel or cells to be processed. The identification information of the processing target may be obtained by optically reading a barcode or the like, or may be manually input by the user or selected from a list. This configuration makes it possible to detach cells according to the different conditions of each sample.

[0049] (Program for implementing cell processing method) The present invention can also be realized by executing the following process. That is, the present invention can be realized by supplying software (programs) that realize one or more functions of the various embodiments described above to a system or device via a network or a storage medium. The present invention can also be realized by a process in which a computer (or a CPU, MPU, or the like) of the system or device reads and executes the program. The computer has one or more processors or circuits, and may include multiple separate computers or a network of multiple separate processors or circuits to read and execute computer-executable instructions.

[0050] In this case, the processor or circuitry may include a central processing unit (CPU), a microprocessing unit (MPU), a graphics processing unit (GPU), an application specific integrated circuit (ASIC), or a field programmable gateway (FPGA), a digital signal processor (DSP), a data flow processor (DFP), or a neural processing unit (NPU).

[0051] The disclosure of the present embodiment includes the following methods and configurations.

[0052] (Method 1) A cell processing method for detaching cells adhered to a culture surface of a culture vessel from the culture vessel using a cell processing system including a vibration unit that generates vibrations, comprising: a heating step of causing the vibration part to vibrate in a first mode while the vibration part and the vibration transmitting material are in contact with each other, thereby increasing the temperature of the vibration transmitting material; a detachment step of detaching the cells from the culture vessel after the start of the heating step by causing the vibration unit to generate vibration in a second mode different from the first mode while the vibration transmission material and the culture vessel are in contact with each other; A cell treatment method comprising the steps of:

[0053] (Method 2) The cell processing method described in Method 1, wherein the vibrations of the first mode and the second mode are vibrations in the ultrasonic band.

[0054] (Method 3) The cell processing method described in Method 1 or 2, wherein the first mode of vibration is a vibration having a higher frequency than the second mode of vibration.

[0055] (Method 4) The cell processing method described in any one of Methods 1 to 3, characterized in that the vibration unit has a vibrator including a piezoelectric material and a vibration plate, and generates vibrations in the first mode and the second mode by applying a drive voltage to the vibrator.

[0056] (Method 5) 5. The cell processing method according to any one of Methods 1 to 4, wherein the vibration of the first mode is a vibration in which the amplitudes of a plurality of different portions on the vibration surface of the vibration unit are maximized.

[0057] (Method 6) 6. A cell treatment method according to any one of Methods 1 to 5, characterized in that there is at least a period of time during which the heating step and the detachment step are carried out simultaneously.

[0058] (Method 7) 7. The cell processing method according to any one of Methods 1 to 6, wherein after the start of the detachment process, the vibration unit is caused to generate vibration of the first mode.

[0059] (Method 8) The cell treatment method described in any one of Methods 1 to 7, wherein the second mode vibration is a square wave or a triangular wave.

[0060] (Method 9) A cell processing method described in any one of methods 1 to 8, characterized in that, in at least one of the heating process and the detachment process, when the temperature of the vibration transmitting material falls outside a predetermined range, the vibration frequency or amplitude of the vibration unit is changed.

[0061] (Method 10) The cell treatment method according to any one of Methods 1 to 9, wherein the vibration transmission material is a solid or liquid.

[0062] (Method 11) The cell treatment method described in Method 10, wherein the vibration transmission material comprises silicone rubber or water.

[0063] (Method 12) A program for causing a computer to execute the cell processing method according to any one of Methods 1 to 11.

[0064] (Configuration 13) A cell treatment device for detaching cells adhered to a culture surface of a culture vessel from the culture vessel, comprising: A vibration unit that generates vibrations and a control unit that controls the vibrations of the vibration unit, The control unit is a vibration unit that is in contact with a vibration transmitting material and that vibrates the vibration unit in a first mode to transmit the vibration to the vibration transmitting material and increase the temperature of the vibration transmitting material; A cell processing device characterized by detaching the cells from the culture vessel by vibrating the vibration unit in a second mode different from the first mode while the vibration transmission material and the culture vessel are in contact with each other.

[0065] (Configuration 14) 14. The cell processing device according to claim 13, wherein the first mode is a vibration having a higher frequency than the second mode.

[0066] (Configuration 15) Further comprising a memory unit that stores processing conditions associated with at least one of the cell type, the culture conditions of the cell, and the type of the culture vessel; 15. The cell processing device according to configuration 13 or 14, wherein the control unit controls the vibration of the vibration unit in accordance with the processing conditions.

[0067] (Configuration 16) A measuring unit for measuring a temperature of the vibration transmitting material; and 16. The cell processing device according to any one of configurations 13 to 15, further comprising an alarm unit that notifies the user by at least one of light, sound, and text display when the temperature measured by the measurement unit reaches a predetermined temperature.

[0068] (Configuration 17) A cell processing system for detaching cells adhered to a culture surface of a culture vessel from the culture vessel, comprising: A vibration unit that generates vibrations and a control unit that controls the vibrations of the vibration unit, The control unit is a vibration unit that is in contact with a vibration transmitting material and that vibrates the vibration unit in a first mode to transmit the vibration to the vibration transmitting material and increase the temperature of the vibration transmitting material; A cell processing system characterized by detaching the cells from the culture vessel by vibrating the vibration unit in a second mode different from the first mode while the vibration transmission material and the culture vessel are in contact with each other.

[0069] (Configuration 18) 18. The cell processing system of claim 17, wherein the first mode is a vibration having a higher frequency than the second mode.

[0070] (Configuration 19) Further comprising a memory unit that stores processing conditions associated with at least one of the cell type, the culture conditions of the cell, and the type of the culture vessel; 19. The cell processing system according to configuration 17 or 18, wherein the control unit controls the vibration of the vibration unit in accordance with the processing conditions.

[0071] (Configuration 20) A measuring unit for measuring a temperature of the vibration transmitting material; and 20. A cell processing system according to any one of configurations 17 to 19, characterized in that it has an alarm unit that notifies by at least one of light, sound, and text display when the temperature measured by the measurement unit reaches a predetermined temperature. [Explanation of symbols]

[0072] 1. Cell processing equipment 3 cells 8 Culture solution 100 culture vessels 101 Case 102 Vibration plate 107 Vibration part 117 Piezoelectric element 129 Vibration Transmission Materials

Claims

1. A cell processing method for detaching cells adhered to a culture surface of a culture vessel from the culture vessel using a cell processing system including a vibration unit that generates vibrations, comprising: a heating step of generating a vibration of a first mode in the vibration part while the vibration part and the vibration transmitting material are in contact with each other, thereby increasing the temperature of the vibration transmitting material; a detachment step of detaching the cells from the culture vessel after the start of the heating step by causing the vibration unit to generate vibration in a second mode different from the first mode while the vibration transmission material and the culture vessel are in contact with each other; A cell treatment method comprising the steps of:

2. 2. The cell treatment method according to claim 1, wherein the vibrations of the first mode and the second mode are vibrations in an ultrasonic band.

3. 2. The cell treatment method according to claim 1, wherein the vibration of the first mode is a vibration of a higher frequency than the vibration of the second mode.

4. The cell processing method according to claim 1, characterized in that the vibration unit has a vibrator containing a piezoelectric material and a vibration plate, and generates vibrations in the first mode and the second mode by applying a drive voltage to the vibrator.

5. 2. The cell treatment method according to claim 1, wherein the vibration of the first mode is a vibration in which the amplitudes of a plurality of different portions on the vibration surface of the vibration section are maximized.

6. The cell treatment method according to claim 1 , further comprising at least a period during which the heating step and the detachment step are carried out simultaneously.

7. 2. The cell treatment method according to claim 1, wherein the vibration unit generates the vibration of the first mode after the start of the detachment step.

8. The cell treatment method according to claim 1 , wherein the vibration of the second mode is a square wave or a triangular wave.

9. The cell treatment method according to claim 1, characterized in that, in at least one of the heating step and the detachment step, if the temperature of the vibration transmitting material falls outside a predetermined range, the vibration frequency or amplitude of the vibration unit is changed.

10. The cell treatment method according to claim 1 , wherein the vibration transmitting material is a solid or a liquid.

11. The cell treatment method according to claim 10, wherein the vibration transmitting material contains silicone rubber or water.

12. A program for causing a computer to execute the cell processing method according to any one of claims 1 to 11.

13. A cell treatment device for detaching cells adhered to a culture surface of a culture vessel from the culture vessel, comprising: A vibration unit that generates vibrations and a control unit that controls the vibrations of the vibration unit, The control unit is a vibration unit that is in contact with a vibration transmitting material and that vibrates the vibration unit in a first mode to transmit the vibration to the vibration transmitting material and increase the temperature of the vibration transmitting material; A cell processing device characterized in that the cells are detached from the culture vessel by vibrating the vibration unit in a second mode different from the first mode while the vibration transmission material and the culture vessel are in contact with each other.

14. The cell processing apparatus according to claim 13, wherein the first mode is a vibration having a higher frequency than the second mode.

15. Further comprising a memory unit that stores processing conditions associated with at least one of the cell type, the culture conditions of the cell, and the type of the culture vessel; The cell processing apparatus according to claim 13 , wherein the control unit controls the vibration of the vibration unit in accordance with the processing conditions.

16. A measuring unit for measuring a temperature of the vibration transmitting material; and The cell processing device according to claim 13, further comprising an alarm unit that notifies the user by at least one of light, sound, and text display when the temperature measured by the measurement unit reaches a predetermined temperature.

17. A cell processing system for detaching cells adhered to a culture surface of a culture vessel from the culture vessel, comprising: A vibration unit that generates vibrations and a control unit that controls the vibrations of the vibration unit, The control unit is a vibration unit that is in contact with a vibration transmitting material and that vibrates the vibration unit in a first mode to transmit the vibration to the vibration transmitting material and increase the temperature of the vibration transmitting material; A cell processing system characterized by detaching the cells from the culture vessel by vibrating the vibration unit in a second mode different from the first mode while the vibration transmission material and the culture vessel are in contact.

18. 18. The cell processing system of claim 17, wherein the first mode is a vibration having a higher frequency than the second mode.

19. Further comprising a memory unit that stores processing conditions associated with at least one of the cell type, the culture conditions of the cell, and the type of the culture vessel; The cell processing system according to claim 17 , wherein the control unit controls the vibration of the vibration unit in accordance with the processing conditions.

20. A measuring unit for measuring a temperature of the vibration transmitting material; and 18. The cell processing system according to claim 17, further comprising an alarm unit that notifies the user by at least one of light, sound, and text display when the temperature measured by the measurement unit reaches a predetermined temperature.

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