WO2021033722A1 - Method for producing habituated cells, method for managing habituated cells, device for managing habituated cells, and computer program - Google Patents

Abstract

A method for producing habituated cells, comprising: obtaining two-dimensionally cultured pluripotent stem cells from a supplier; subjecting the obtained pluripotent stem cells to a habituation treatment for enabling the transfer from two-dimensional culture to suspension culture; obtaining pluripotent stem cells that have been habituated for suspension culture by the preceding habituation treatment; and providing the habituated pluripotent stem cells to a specified customer on demand from the customer.

Description

Adaptation cell preparation method, acclimation cell management method, acclimation cell management device, and computer program

The present disclosure relates to a method for producing acclimated cells, a method for managing acclimated cells, an acclimatized cell management device, and a computer program.

Embryonic stem cells (ES cells) and inducible pluripotent stem cells (iPS cells) are mainly known as pluripotent stem cells. These pluripotent stem cells retain the ability to differentiate into other cell lines and can differentiate into cells of the desired stage under various conditions. Taking advantage of this property, the use of pluripotent stem cells is increasing in many fields such as cell differentiation research, disease treatment application, and drug screening.

By the way, for culturing these pluripotent stem cells, a cell scaffold material formed of an adhesive substrate material is used, but in order to obtain a larger amount in a smaller space, a suspension culture that does not require a cell scaffold material is used. It has been proposed (eg, WO2011 / 058558, US901801, etc.).

However, since pluripotent stem cells are originally adhesive cells, they tend to be difficult to apply immediately to suspension culture. For this reason, it takes time and labor to start the acclimatization process after a certain amount of pluripotent stem cells are required.
An object of the present disclosure is to provide a method for producing acclimated cells to obtain pluripotent stem cells acclimated to suspension culture while saving time and effort. In addition, it is necessary to appropriately manage the acclimation process by acquiring various information such as information on the pluripotent stem cells to be subjected to the acclimation process, information on the acclimated pluripotent stem cells, and information on the acclimation process itself. In addition, acclimation cell production methods, acclimation cell management methods, acclimation cell management devices, and computer programs that can provide additional value in providing acclimated pluripotent stem cells to customers. Another purpose is to provide.

The present disclosure provides the following aspects.
The method for producing acclimated cells according to one aspect of the present disclosure enables the reception of flat-cultured pluripotent stem cells from a supplier and the transition of the received pluripotent stem cells from flat culture to suspension culture. To obtain pluripotent stem cells acclimated to suspension culture by the acclimation treatment, and to specify the obtained acclimated pluripotent stem cells according to the request from the provider. Including providing to the destination.

Further, in the method for producing acclimated cells according to one aspect, the pluripotent stem cells received by the acclimation treatment are subjected to primary acclimation culture in a primary acclimation culture vessel filled with a primary acclimation medium, and the primary acclimation has been performed. The primary acclimation treatment to obtain pluripotent stem cells and the suspension culture of the primary acclimated pluripotent stem cells in a suspension culture container filled with a secondary acclimation medium were performed, and the secondary acclimated polyplasia was acclimated to the suspension culture. Includes a secondary acclimation process to obtain capable stem cells.

In addition, the method for producing acclimated cells according to one embodiment includes initial cell information regarding the pluripotent stem cells before being subjected to the acclimation treatment, primary acclimation processing information regarding the primary acclimation treatment, and secondary acclimation treatment. Selected from the group consisting of acclimation processing information, primary acclimated pluripotent stem cell information on the primary acclimated pluripotent stem cell, and secondary acclimated pluripotent stem cell information on the secondary acclimated pluripotent stem cell. Further including obtaining at least one piece of information.

Further, in the method for producing an acclimated cell according to one aspect, the initial cell information includes at least information on the cell type of the pluripotent stem cell before being subjected to the acclimation treatment.

Further, in the method for producing acclimated cells according to one aspect, the primary acclimation treatment information includes at least information on the primary acclimation medium and information on culture conditions related to the primary acclimation treatment.

Further, in the method for producing acclimated cells according to one aspect, the secondary acclimation treatment information includes at least information on the secondary acclimation medium and information on suspension culture conditions related to the secondary acclimation treatment.

Further, in the method for producing acclimated cells according to one aspect, the primary conditioned pluripotent stem cell information includes at least information on the number of cells of the primary conditioned pluripotent stem cells and the characteristics of cell morphology, and the secondary The conditioned pluripotent stem cell information includes at least information on the cell number of the secondary conditioned pluripotent stem cells and the characteristics of cell morphology.

In addition, the method for producing acclimated cells according to one embodiment includes the initial cell information, the primary acclimation processing information, the secondary acclimation processing information, the primary acclimated pluripotent stem cell information, and the secondary acclimated pluripotency. Further comprising providing the provider with at least one obtained information selected from the group consisting of stem cell information.

Further, in the method for producing acclimated cells according to one aspect, the acclimated pluripotent stem cells are provided to the donor in a suspended culture form.

The acclimation cell management method according to one aspect of the present disclosure is a acclimation cell management method executed by at least one processor that executes a computer-readable instruction, and the at least one processor executes the instruction. Thereby, the initial cell information regarding the planar cultured pluripotent stem cells is acquired and stored, and the acclimatization processing information regarding the acclimatization process for enabling the transition from the planar culture to the suspension culture for the pluripotent stem cells is performed. Is acquired and stored, and the acclimatized pluripotent stem cell information regarding the acclimatized pluripotent stem cells generated by the acclimatization process is acquired and stored.

Further, in the acclimated cell management method according to one embodiment, the at least one processor executes the instruction to further provide the acclimated pluripotent stem cell to the destination to which the acclimated pluripotent stem cell is provided. The donor request information regarding the request for the sex stem cell is acquired and stored in advance, and the donor request information is compared with the acclimatized pluripotent stem cell information to determine whether or not there is a difference between the two.

Further, in the acclimation cell management method according to one aspect, the at least one processor further obtains the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information by executing the instruction. At least one piece of information selected from the group is transmitted to the destination server device to which the acclimatized pluripotent stem cells are provided via a communication line.

Further, in the acclimation cell management method according to one aspect, the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information are acquired from the acclimation processing device that executes the acclimation processing via a communication line. Will be done.

Further, in the acclimation cell management method according to one aspect, the provision destination request information is acquired from the provision destination server device of the provision destination via a communication line.

Further, in the acclimation cell management method according to one aspect, in the acclimation treatment, the pluripotent stem cells are subjected to primary acclimation culture in a primary acclimation culture vessel filled with a primary acclimation medium, and the primary acclimated pluripotency is performed. The primary acclimation treatment for obtaining sex stem cells and the suspension culture of the primary acclimated pluripotent stem cells in a suspension culture container filled with a secondary acclimation medium are performed, and the secondary acclimated pluripotency acclimated to the suspension culture is performed. The acclimation processing information includes the primary acclimation processing information regarding the primary acclimation process and the secondary acclimation processing information regarding the secondary acclimation process, and includes the secondary acclimation process for obtaining sex stem cells. The stem cell information includes the primary acclimated pluripotent stem cell information regarding the primary conditioned pluripotent stem cell and the secondary conditioned pluripotent stem cell information regarding the secondary conditioned pluripotent stem cell.

Further, in the acclimated cell management method according to one aspect, the initial cell information includes at least information on the cell type of the pluripotent stem cell.

Further, in the acclimation cell management method according to one aspect, the primary acclimation treatment information includes at least information on the primary acclimation medium and information on culture conditions related to the primary acclimation treatment.

Further, in the acclimation cell management method according to one aspect, the secondary acclimation processing information includes at least information on the secondary acclimation medium and information on suspension culture information on the secondary acclimation treatment.

Further, in the conditioned cell management method according to one aspect, the primary conditioned pluripotent stem cell information includes at least information on the number of cells of the primary conditioned pluripotent stem cell and the characteristics of cell morphology, and the secondary The conditioned pluripotent stem cell information includes at least information on the cell number of the secondary conditioned pluripotent stem cells and the characteristics of cell morphology.

The acclimated cell management apparatus according to one aspect of the present disclosure includes one or a plurality of processors, and the processors acquire and store initial cell information regarding plane-cultured pluripotent stem cells, and the pluripotent stem cells. On the other hand, the acclimation process information related to the acclimation process for performing the acclimation process to enable the transition from the planar culture to the suspension culture is acquired and stored, and the acclimated pluripotent stem cells generated by the acclimatization process have been acclimatized. The at least one processor is made to function so as to acquire and store pluripotent stem cell information.

In addition, the acclimated cell management device according to one aspect acquires and stores in advance destination request information regarding a request for the acclimated pluripotent stem cell of the destination to which the acclimated pluripotent stem cell is provided. The at least one processor is further functionalized to compare the destination request information with the acclimatized pluripotent stem cell information to determine if there is a difference between the two.

In addition, the acclimatized cell management device according to one aspect transmits at least one information selected from the group consisting of the initial cell information, the acclimatization processing information, and the acclimatized pluripotent stem cell information via a communication line. The at least one processor is further functionalized to transmit the conditioned pluripotent stem cells to a destination server device to which the cells are provided.

Further, in the acclimation cell management device according to one aspect, the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information are acquired from the acclimation processing device that executes the acclimation processing via a communication line. Will be done.

Further, in the acclimatized cell management device according to one aspect, the provision destination request information is acquired from the provision destination server device of the provision destination via a communication line.

The computer program according to one aspect of the present disclosure acquires and stores initial cell information about pluripotent stem cells cultured in a plane by being executed by at least one processor, and planes the pluripotent stem cells. Performs acclimation processing to enable the transition from culture to suspension culture Acquires and stores acclimation processing information related to the acclimation process, and acclimatized pluripotent stem cells related to the acclimated pluripotent stem cells generated by the acclimation process. The at least one processor is made to function so as to acquire and store information.

In addition, the computer program according to one aspect acquires, stores, and stores in advance the destination request information regarding the request for the conditioned pluripotent stem cell of the destination to which the conditioned pluripotent stem cell is provided, and provides the provided information. The at least one processor is further functionalized to compare the prior demand information with the acclimated pluripotent stem cell information to determine if there is a difference between the two.

Further, the computer program according to one aspect obtains at least one piece of information selected from the group consisting of the initial cell information, the acclimatization processing information, and the acclimatized pluripotent stem cell information via a communication line. The at least one processor is further functionalized such that the conditioned pluripotent stem cells transmit to the destination server device of the destination to which they are provided.

Further, in the computer program according to one aspect, the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information are acquired from the acclimation processing device that executes the acclimation processing via a communication line. ..

Further, in the computer program according to one aspect, the provision destination request information is acquired from the provision destination server device of the provision destination via a communication line.

According to the present disclosure, it is possible to provide a method for producing acclimated cells to obtain pluripotent stem cells acclimated to suspension culture while saving time and effort. Further, according to the present disclosure, the acclimatization process is performed by acquiring various information such as information on the pluripotent stem cells to be subjected to the acclimatization process, information on the acclimatized pluripotent stem cells, and information on the acclimatization process itself. Providing acclimation cell management methods, acclimation cell management devices, and computer programs that can be properly managed and also provide additional value in providing acclimated pluripotent stem cells to customers. can do.

It is a block diagram which shows an example of the structure of the management system including the acclimatization cell management apparatus which concerns on one Embodiment. It is a block diagram which shows typically an example of the function of the acclimation cell management apparatus shown in FIG. It is a block diagram which shows typically an example of the function of the acclimatization processing apparatus shown in FIG.

In the present specification, the numerical range indicated by using "-" indicates a range including the numerical values before and after "-" as the minimum value and the maximum value, respectively. In the present specification, the content of each component in the composition is the total amount of the plurality of substances present in the composition unless otherwise specified, when a plurality of substances corresponding to each component are present in the composition. Means. The term "process" is included in the term not only in an independent process but also in the case where the desired action of the process is achieved even if it cannot be clearly distinguished from other processes.

The method for producing acclimated cells according to one aspect of the present disclosure is to receive the pluripotent stem cells cultured in a plane from a supplier (hereinafter referred to as a receiving step), and to suspend the received pluripotent stem cells from a plane culture. Performing an acclimatization treatment to enable the transition to (hereinafter referred to as an acclimatization step), obtaining pluripotent stem cells acclimatized to suspension culture by the acclimatization treatment, and the obtained pluripotent pluripotency It includes providing stem cells to a designated donation destination (hereinafter referred to as a donation step) in response to a request from the donation destination.
According to these methods for producing acclimated cells of the present disclosure, pluripotent stem cells acclimated to suspension culture can be obtained while saving time and effort.

To further explain this, according to this method for producing acclimated cells, the pluripotent stem cells cultured in a plane are obtained from a supplier. The acclimated pluripotent stem cells obtained by subjecting the obtained pluripotent stem cells to a specific acclimation treatment are provided to a designated donor in response to a request from the donor. Therefore, the donor can obtain the pluripotent stem cells that have been acclimatized to the suspension culture when they are requested without the need for acclimatization treatment. Here, the supply source and the supply destination may be the same or different. In the present specification, the description of "provider" may be read as "supply source", and in some cases, may be read as both "provider" and "supply source". The acclimation process is carried out at a place, institution, etc. different from the supply source and the supply destination. Hereinafter, in the present specification, a place, an institution, etc. that performs habituation processing may be referred to as a "habituation executing agency" for convenience. In the present specification, the culture medium may be simply referred to as "medium".
Hereinafter, the method for producing the acclimated cells will be described.

1. 1. In the process of receiving the acclimated cell preparation method , the flat-cultured pluripotent stem cells are received from the supplier. In the present specification, the “pluripotent stem cell” is not limited as long as it is an undifferentiated cell having the ability to differentiate into a plurality of cells and self-renewal ability, and is an embryonic stem cell (ES cell) or an inducible pluripotent stem cell. (IPS cells), mesenchymal stem cells (MSCs), Muse cells and the like can be mentioned, and among them, ES cells and iPS cells capable of differentiating into all three embryonic lineages of endometrial, mesenchymal and ectodermal can be used. .. The origin of these pluripotent stem cells is not particularly limited, and examples thereof include humans, monkeys, mice, rats, pigs, and dogs.

The pluripotent stem cells obtained in the receiving step are plane-cultured. Here, the planar culture is also referred to as two-dimensional culture in some cases, and refers to a culture method in which cells are adhered and cultured on the culture surface of a culture vessel having a flat surface. The plane that becomes the culture surface of the culture vessel is a cell of a cell-affinitive polymer, a cell layer of a feeder cell, an extracellular matrix such as vitronectin, fibronectin, collagen, Matrigel (registered trademark, hereinafter omitted), or a partial peptide thereof. It may be provided as a scaffolding material. The medium used for the planar culture can be appropriately selected depending on the type of pluripotent stem cells to be targeted, and examples thereof include DMEM or DMEM-F12 containing 10% (v / v) FBS. The culture conditions for the planar culture may be any conditions normally applied to the target pluripotent stem cells, and can be, for example, a temperature condition of 37 ° C. and a 10% CO ₂ environment. The culture conditions can be appropriately changed according to the state of the cells as long as the differentiation stage of the cells does not change.

As the receiving method, generally, any form can be provided as long as the cells can be transferred, and examples thereof include a liquid culture form and a cryopreservation form. Pluripotent stem cells may be transferred from the source to the acclimatization agency. For the transfer, a transfer form such as a constant temperature transfer, a refrigerated transfer, a frozen transfer, or the like may be appropriately selected according to the form of provision from the supplier.

In the acclimation step, the received pluripotent stem cells are acclimated to enable the transition from planar culture to suspension culture. As used herein, the term "suspended culture" refers to a culture method in which cells are suspended and cultured in a medium rather than being adhered to a flat surface. In other words, in the present specification, the acclimatization treatment refers to a culture medium in which pluripotent stem cells adapted to a static growth environment that grows by adhering to a plane that becomes a two-dimensional environment are used as a three-dimensional environment. It is a process of gradually "acclimating" cells to a floating environment in which cells are dispersed and grow in the system. In the acclimation step, the cells may be dispersed in the medium as a single cell, a plurality of cells may accumulate to form a spheroid, and the cells adhere to microcarriers such as microbeads to which they can adhere. It may be in a state or may have periods of these different states during the acclimatization process. A plurality of media may be used for the acclimation treatment, and the culture conditions such as the culture temperature, pH, and stirring of the medium may be changed. In the culture in the acclimation step, cells can be maintained and increased by repeating the passage at a predetermined timing so as to maintain an appropriate number of cells according to the state of the cells. The culture can be carried out under a temperature condition of, for example, 37 ° C. and in a 10% CO _{2 environment.}
By going through the acclimation step, pluripotent stem cells acclimated to suspension culture can be obtained.

In the acclimatization treatment, the received pluripotent stem cells are further subjected to primary acclimatization culture in a primary acclimatization culture vessel filled with a primary acclimatization medium to obtain primary acclimatized pluripotent stem cells (hereinafter, primary acclimatization treatment). ), The primary acclimated pluripotent stem cells are subjected to suspension culture in a suspension culture container filled with a secondary acclimation medium (hereinafter referred to as secondary acclimation treatment), and the secondary acclimation medium is used for suspension culture. It can include obtaining secondary conditioned pluripotent stem cells.
By performing the acclimation treatment in two stages in which the medium and the culture vessel are different, pluripotent stem cells acclimated to the suspension culture can be more reliably obtained.

For the primary acclimation treatment, a primary acclimation medium and a primary acclimation culture container are used. The primary acclimation medium and the primary acclimation culture vessel used in the primary acclimation treatment may be at least one different from the medium or culture vessel used in the plane culture before receiving the pluripotent stem cells. As a result, pluripotent stem cells can be provided in a culture environment different from that of the planar culture before receipt. In the primary acclimation treatment, if at least one of the medium and the culture vessel is different from the culture conditions of the plane culture or the secondary acclimation treatment before receiving the cells, the plane culture stage and the floating under the conditions different from the secondary acclimation treatment. At least one stage with the culture stage can be included, and both the plane culture stage and the suspension culture stage can be included. By making the primary acclimation process a multi-step primary acclimation process consisting of two stages, a plane culture stage and a suspension culture stage, it is possible to perform cell acclimation gently with less stress due to acclimation, and the cells float more efficiently. Pluripotent stem cells acclimated to culture can be obtained.

The culture conditions at the plane culture stage in the primary acclimatization treatment may be the same as or different from the plane culture performed on the received pluripotent stem cells. Examples of the culture conditions referred to here include the medium to be used, the culture vessel, temperature, pH and the like. In the planar culture step in the primary acclimation treatment, extracellular matrix such as Matrigel and cell scaffold material such as mouse embryonic fibroblast (MEF) may be used on the culture surface, and Matrigel having no heterologous component may be used. Can be used.

For the planar culture step in the primary acclimation treatment, a medium different from the planar culture performed on the received pluripotent stem cells can be used. In the plane culture stage in the primary acclimation treatment, for example, a medium containing DMEM-F12 containing human basic fibroblast growth factor (bFGF), TGFβ1, GABA, bovine serum albumin (BSA), cholesterol, etc. is used for the plane culture stage. It can be used as a medium. As the culture vessel, for example, a dish coated with Matrigel as a cell scaffolding material can be used. The plane culture step can be completed when the number of cells reaches ^{a predetermined number, for example, about 1 × 10 6} to 3 × 10 ^6. When the suspension culture stage is performed following the plane culture stage, the cells in the plane culture can be recovered by enzyme treatment such as trypsin and can be transferred to the suspension culture stage.

In the suspension culture stage in the primary acclimatization treatment, culture conditions that facilitate suspension culture of pluripotent stem cells, for example, selection of a medium having a composition that facilitates suspension culture of pluripotent stem cells, removal of cell scaffold material, a combination thereof, etc. are selected. can do. As a result, the pluripotent stem cells that have been cultivated in a plane are subjected to suspension culture. Examples of the culture medium for the suspension culture stage having a composition that facilitates suspension culture of pluripotent stem cells include 1 to 50% (v / v) or 5 to 30% (v / v) knockout serum substitute, 1 to 150 ng /. Examples thereof include a medium containing mL of basic fibroblast growth factor (bFGF), L-glutamine, a reducing agent, and the like. The suspension culture step in the primary acclimation treatment can be static suspension culture. Examples of the culture vessel used in the static suspension culture step include a dish having no cell scaffolding material such as a matrix such as Matrigel or a mouse embryonic fibroblast layer.

The suspension culture step in the primary acclimation process can be continued until the pluripotent stem cells enter the logarithmic growth phase in which they proliferate in an undifferentiated state. Although it depends on the state of cells, it can generally be about 1 to 2 months. Pellets of pluripotent stem cells in suspension culture or isolated pluripotent stem cell clusters can be dissociated by pipetting the cells up and down in a small amount (eg, 0.2-1 mL) of medium. it can. As a result, pluripotent stem cells in the suspension culture stage can be efficiently obtained in large quantities. When it is confirmed that the pluripotent stem cells have entered the logarithmic growth phase in the undifferentiated state, the primary acclimation treatment step is terminated and the process proceeds to the secondary acclimation step. By the primary acclimation step, primary acclimated pluripotent stem cells can be obtained.

In the secondary acclimation step, the primary acclimated pluripotent stem cells are suspended-cultured in a suspension culture container filled with a secondary acclimation medium. The medium for secondary acclimation used in the secondary acclimation step can be different from the medium in the suspension culture stage in the primary acclimation step. Specifically, it is possible to select a medium having a composition that enhances the tendency for suspension culture to be more likely for the primary conditioned pluripotent stem cells.

Examples of the medium for secondary acclimation include a medium containing a protease inhibitor.
As the protease inhibitor, those described later can be used. Mediums containing protease inhibitors can contain naturally occurring components. As the medium containing the protease inhibitor, a limited medium containing the protease inhibitor may be used. As used herein, the term "limited medium" means a culture medium that is artificial and whose components are all known. The medium can be an aqueous medium containing a combination of substances such as salts, nutrients, minerals, vitamins, antibiotics, amino acids, nucleic acids, and proteins. For example, various additives described in the specification can be added to a synthetic tissue culture medium such as DMEM-F12.

Examples of the protease inhibitor include a reversible protease inhibitor and an irreversible protease inhibitor, and a reversible protease inhibitor and an irreversible protease inhibitor of serine protease. Reversible inhibitors of serine proteases include phenylmethylsulfonylfluoride (PMSF), GSK3β inhibitors, aldehyde-CHO, arylketone- _{CO-aryl, trifluoromethylketone-COCF 3} , ketocarboxylic acid-COCOOH and the like. .. For example, the concentration of PMSF used can be 0.01 mM to 1 mM in the medium. Irreversible inhibitors of serine proteases include trypsin and trypsin-like serine proteases, including tosyl-L-lysyl-chloromethanehydrochloride (TLCK). TLCK (CAS 4238-41-9) is an irreversible inhibitor of trypsin and trypsin-like serine proteases. The concentration of TLCK used can be 0.05 μM to 1000 μM in the medium, for example, 5 μM to 100 μM.

The medium for secondary acclimation can further contain a GSK3β inhibitor such as CHIR99021, a WNT3A polypeptide, a stabilizer for WNT3A, a leukemia inhibitory factor (LIF), an IL6RIL6 chimera and the like. The medium for secondary acclimation can be free of ERK1 / 2 inhibitors, JNK inhibitors, p38 inhibitors, basic fibroblast growth factor (bFGF) and the like. The secondary acclimation medium can be serum-free and can include 1-50% (v / v) or 5-30% (v / v) knockout serum substitutes. Examples of such a secondary acclimation medium include the limited culture medium described in International Publication No. 2018/015954 (US2019 / 0284526A).
Examples of the medium for secondary acclimation include Essential 8 (Thermo Fisher), mTeSR1 (STEMCELL Technologies), StemFit series (Takara Bio Inc.), StemFlex (Thermo Fisher Scientific), and the like, in addition to those described above. it can.

The suspension culture in the secondary acclimation step can be a dynamic suspension culture in order to expand the number of cells more efficiently. The suspension culture container used for dynamic suspension culture is particularly limited as long as it has a volume and equipment capable of dispersing the cells in culture in the medium at an appropriate speed, shearing force, and dispersion degree. However, for example, Spinna Plasco and the like can be mentioned.

The medium used in the suspension culture stage of the primary acclimation treatment can be switched to the secondary acclimation medium in several stages of two or more stages so as to gradually increase the proportion of the secondary acclimation medium. The culture vessel may be changed from a culture vessel suitable for static suspension culture to a culture vessel suitable for dynamic suspension culture at a timing according to the cell growth state in accordance with the change in the composition of the medium.
As a result, the primary conditioned cells can be gently applied to the secondary conditioned environment, and as a result, the number of cells can be efficiently expanded.

The secondary acclimation step may be continued while grasping the state of the cells while exchanging the medium and confirming that the cells are in an undifferentiated state. Pellets of pluripotent stem cells in suspension culture or isolated pluripotent stem cell clusters can be dissociated by pipetting the cells up and down in a small amount (eg, 0.2-1 mL) of medium. it can. As a result, pluripotent stem cells in suspension culture can be efficiently obtained in large quantities. After several weeks of secondary acclimation, secondary acclimated pluripotent stem cells acclimated to suspension culture can be obtained. The undifferentiated state of the secondary conditioned pluripotent stem cells can be confirmed by appearance, cell surface antigen, expression marker and the like. Being undifferentiated is a known feature that confirms the undifferentiated state of pluripotent stem cells, such as at least 2 of markers such as OCT4, Nanog, Tra-1-60, Tra-1-81, SSEA4, SOX2. It can be confirmed in one or more expression states.

In the donation process, the obtained acclimatized pluripotent stem cells are provided by the acclimatization executing agency to the designated donor in response to a request from the donor. As mentioned above, the provider may be the same as or different from the supplier, and therefore the offer may be in response to a request from the supplier or a request from the supplier. May be good.

Examples of the form of provision include the following forms:
(1) A form in which acclimatized pluripotent stem cells derived from pluripotent stem cells received from the supplier are provided to the supplier in response to a request from the supplier, that is, when the supplier requires it.
(2) The conditioned pluripotent stem cells derived from the pluripotent stem cells received from the supplier are sent to a different destination from the supplier in response to a request from the supplier, that is, when the supplier requires it. Form to provide,
(3) A form in which the conditioned pluripotent stem cells derived from the pluripotent stem cells received from the supplier are provided to the donor in response to a request from the donor, that is, when the donor requires it.

The acclimatized pluripotent stem cells provided to the recipient may be provided in a suspension culture form, or may be isolated from the suspension culture form and provided, for example, in a cryopreserved form. When provided in the suspension culture form, the acclimatized pluripotent stem cells can be collected from the suspension culture system and then filled in a supply container together with a predetermined medium. The container for provision may be a hard container such as a bottle or a soft container such as a bag. Examples of such a container for provision include a bioreactor for cell culture. The bioreactor for cell culture may be a reusable one, or may be a so-called single-use bioreactor. Pluripotent stem cells can be transferred from the acclimatization executing agency to the delivery destination. For the transfer, a transfer form such as a constant temperature transfer, a refrigerated transfer, a frozen transfer, or the like may be appropriately selected according to the supply form.

As described above, according to this method for producing acclimated cells, the supplier or the provider using the pluripotent stem cells acclimated to the suspended cells does not have to take the trouble of carrying out the steps from the planar culture to the suspended culture by themselves. Pluripotent stem cells acclimated to can be obtained.

In other words, pluripotent stem cells acclimatized to floating cells can be maintained like a "fish cage" and easily prepared with the required number of cells when needed. For example, when the source or destination of cells is the customer, it is not necessary for the customer to prepare the acclimated pluripotent stem cells, and the number of cells acclimated to the suspension culture is increased when the customer needs it. It is possible to provide pluripotent stem cells.

Since the obtained pluripotent stem cells acclimated to the suspension culture can maintain undifferentiated ability and proliferate efficiently, they can be used in various modes as convenient pluripotent stem cells for the recipient. be able to. Examples of such utilization include utilizing the undifferentiated ability of the acclimatized pluripotent stem cells to induce differentiation into desired cells, and using the cells for screening of drugs and the like.

In this method for producing acclimated cells, when pluripotent stem cells are received from the supplier, an identification ID for distinguishing them from other pluripotent stem cells such as an identification tag, an individual chip, a process control number, and a barcode is assigned. You may. By assigning the identification ID, individual management can be performed from the receiving process to the acclimatization step and the provision process until the acclimatized pluripotent stem cells are provided to the recipient.

In this method for producing acclimated cells, the acclimation process can be performed while monitoring the number of cells (acquiring information on the number of cells). For example, the primary acclimation culture and the secondary acclimation culture can be performed so as to have a predetermined number of cells. The acclimatized cells can be maintained and proliferated while subcultured. The number of cells can be easily monitored, for example, by considering the doubling time of cells and adjusting the number of cells to a certain number at the time of medium exchange and seeding. In this method for producing acclimated cells, information on the number of cells is used as an example, information on pluripotent stem cells to be subjected to acclimation treatment, information on acclimated pluripotent stem cells (acclimated pluripotent stem cells), and acclimation treatment. Various information related to the acclimatization process such as the information itself can be acquired. Details such as acquisition of various information will be described later.

Although the method for producing acclimated cells according to the present disclosure has been described as the cells received from the supplier as pluripotent stem cells cultured in a plane, the cells received from the supplier are not limited to pluripotent stem cells. Another aspect of the disclosure may include receiving plane-cultured somatic cells from a source. In this embodiment, after receiving the cells from the source, the cells are reprogrammed according to a known method, and the pluripotent stem cells obtained by the reprogramming can be transferred from the planar culture to the suspension culture. It may include performing acclimatization processing for the purpose.

2. 2. Acquisition of various information, etc. Next, details of acquisition of various information related to the acclimatization process, etc. will be described with reference to FIGS. 1 to 3. FIG. 1 is a block diagram showing an example of the configuration of the management system 1 including the acclimation cell management device 100 according to the embodiment. FIG. 2 is a block diagram schematically showing an example of the function of the conditioned cell management device 100 shown in FIG. FIG. 3 is a block diagram schematically showing an example of the function of the acclimation processing device 200 shown in FIG.

It should be noted that, in the present specification, the term application may be referred to as software or program, and is a command to a computer, which is combined so as to obtain a certain result. All you need is.

As shown in FIG. 1, the management system 1 is mainly composed of the acclimation cell management device 100 and the acclimation processing device 200. The acclimation cell management device 100 and the acclimation processing device 200 can transmit and receive various electronic information via a communication line. Further, the acclimatization cell management device 100 may be capable of transmitting and receiving various electronic information via a communication line with one or a plurality of destination server devices 300. In addition, in FIG. 1, as an example, it is assumed that there are three providing destinations, and the providing destination server device 300 (providing destination server device 300A, providing destination server device 300B, each of the three providing destinations). And is connected to the provider server device 300C) via a communication line. Hereinafter, the details of the acclimation cell management device 100 and the acclimation processing device 200 constituting the management system 1 will be described.

2-1. Acclimated cell management device 100
The acclimation cell management device 100 is a terminal device or a server device connected to the acclimation processing device 200 via a communication line. The conditioned cell management device 100 may be one terminal device or server device, or may be composed of a plurality of terminal devices or server devices for the purpose of distributing the load. The acclimatized cell management device 100 as a terminal device or a server device is a central processing unit, a main storage device, an input / output interface, an input device, an auxiliary storage device, which are connected to each other by a data bus and / or a control bus as hardware. , An output device, etc., and can execute various information processing described later.

The central processing unit is called a "CPU", and can perform calculations on instructions and data stored in the main storage device, and store the results of the calculations in the main storage device. Further, the central processing unit can control an input device, an auxiliary storage device, an output device, and the like via an input / output interface. The conditioned cell management device 100 can include one or more central processing units.

The main storage device is called a "memory" and can store commands and data received via an input device, an auxiliary storage device, and an input / output interface, as well as calculation results of a central processing unit. The main storage device may include, without limitation, RAM (random access memory), ROM (read-only memory) and / or flash memory and the like.

The auxiliary storage device is a storage device having a larger capacity than the main storage device. The auxiliary storage device can store instructions and data (computer programs) that constitute a specific application, specifically, an acclimation cell management application, a web browser application, and the like. Further, the auxiliary storage device can transmit these instructions and data (computer program) to the main storage device via the input / output interface by being controlled by the central processing unit. The auxiliary storage device may include, but is not limited to, a magnetic disk device and / or an optical disk device and the like.

The input device is a device that takes in data from the outside, and can include a touch panel, a button, a keyboard, a mouse, and / or a sensor without limitation.

The output device can include, without limitation, a display device, a touch panel and / or a printer device, and the like.

In such a hardware configuration, the central processing unit sequentially loads the instructions and data (computer programs) that make up a specific application stored in the auxiliary storage device into the main storage device, and the loaded instructions and data. Can be calculated. Thereby, the central processing unit controls the output device via the input / output interface, or also via the input / output interface and the communication line, the acclimation processing unit 200, and in some cases one or more destination servers. Various information can be transmitted and received to and from the device 300.

A specific application for acquiring various information related to the acclimatization process (for example, the above-mentioned acclimatization cell management application) is installed in the acclimatization cell management device 100. As a result, the acclimatization cell management device 100 can acquire various information related to the acclimatization process by executing the application.

As shown in FIG. 2, the acclimation cell management device 100 having the above-mentioned hardware configuration can mainly include the first communication unit 110 and the storage unit 120. Further, when the acclimatized cell management device 100 is connected to one or a plurality of destination server devices 300 via a communication line, the determination unit 130 can be further included. Furthermore, when the acclimatization cell management device 100 is a terminal device in particular, the display unit 140 and the first user interface unit 150 can be further included.

The first communication unit 110 is at least one selected from a group consisting of various information including initial cell information, acclimation processing information, and acclimated pluripotent stem cell information from the acclimation processing device 200 via a communication line. The information can be received and the received information can be transmitted to the storage unit 120. The details of the initial cell information, the acclimatization processing information, and the acclimatized pluripotent stem cell information will be described later.

Further, when the acclimatized cell management device 100 is connected to one or a plurality of destination server devices 300 via a communication line, the first communication unit 110 is provided by the provider server device 300 via the communication line. It is possible to receive the request information (delivery destination request information) regarding the (desired) conditioned pluripotent stem cell requested by the destination (customer) and transmit the received provision destination request information to the storage unit 120.

The provider request information includes information on the number of acclimated pluripotent stem cells and information on the degree of acclimation of the acclimated pluripotent stem cells, in addition to identification information on the corporate name, personal name, etc. of the provider. (For example, information such as requesting only the primary acclimation process, requesting both the primary acclimation process and the secondary acclimation process, requesting only a part of the primary acclimation process and the secondary acclimation process, etc.) is included as an example. Can be included as. In addition to this information, the provider request information includes information on culture conditions related to acclimatization (medium, culture vessel, temperature, pH, seeding density, number of subcultures, conditions related to subculture conditions, etc.), and acclimatized. Information on the target site (eg, retina, myocardium, nerve, etc.) of pluripotent stem cells, information on the donation form (eg, suspension culture form, cold storage form, etc.), and usage form of the donation destination (for example, Information on induction of differentiation into desired cells, screening of drugs, etc.) can be included without limitation.

Information on the type of differentiated cell of interest in the case of performing a differentiation-inducing treatment for inducing differentiation of the acclimated pluripotent stem cell into a predetermined cell after the acclimation treatment on the acclimated pluripotent stem cell obtained by the acclimation treatment. , And when the provider requests optional information such as genetic testing, such information can also be included in the provider request information. In addition, when performing a safety confirmation process to carry out a designated safety test on the obtained cells, the provider provides information on safety such as impurity test, sterility / mycoplasma denial test, endotoxin test, etc. When requesting, this information can also be included in the destination request information. Furthermore, when transferring designated cells (acclimated pluripotent stem cells) to the delivery destination, if the delivery destination requests shipping management information such as product status and transport information, the shipping management information is also included in the delivery destination request information. Can be included.

By the way, the above-mentioned communication line can include wired, wireless LAN, the Internet, etc. without limitation.

Further, when the acclimatization cell management device 100 is connected to one or a plurality of destination server devices 300 via a communication line, the first communication unit 110 receives initial cell information and acclimatization processing from the acclimatization processing device 200. Various information including the information and the acclimatized pluripotent stem cell information can be transmitted to the providing destination (the server device 300 of the providing destination). As a result, the recipient (customer) is not only provided with physically acclimatized pluripotent stem cells, but also provides information related to the acclimatization process, especially information related to the acclimatized pluripotent stem cells themselves. It will be possible to receive. Therefore, it is possible to receive the donated pluripotent stem cells that are highly reliable and highly reproducible for the recipient.

The storage unit 120 can store the information received from the first communication unit 110. Here, the storage unit 120 can classify and store various information received from the first communication unit 110 according to the destination, the type of pluripotent stem cell, and the like. In this case, for example, when the pluripotent stem cells received from the supplier are given the identification ID as described above, each of the various information received from the acclimatization processing device 200 by the first communication unit 110 is By so that the identification IDs are associated with each other, the storage unit 120 can manage and store various information for each identification ID. Further, the storage unit 120 can also store various information input via the first user interface unit 150, which will be described later, and information related to various settings.

When the acclimation cell management device 100 is connected to one or a plurality of destination server devices 300 via a communication line, the determination unit 130 is a acclimated pluripotent stem cell stored in the storage unit 120 as described above. It is possible to determine whether or not there is a difference between the two after reading out the destination request information regarding the above and the acclimatized pluripotent stem cell information from the storage unit 120 and comparing the two. Specifically, for example, information on the number of cells of the acclimatized pluripotent stem cells (for example, information of "X") as the destination request information and the acclimatized pluripotent stem cells transmitted from the acclimatization processing apparatus 200. Compared with information on the number of acclimatized pluripotent stem cells as information (for example, information with "Y"), X and Y match (or the difference between X and Y is a predetermined range. If (inside), it is judged that there is no difference between the two, and if X and Y do not match (or the difference between X and Y exceeds a predetermined range), there is a difference between the two. judge.

The comparison between the provider request information and the acclimatized pluripotent stem cell information by the determination unit 130 and the determination of the presence or absence of a difference between the two need not be limited to focusing on the number of cells, and other details are given. It may be replaced with information or executed in combination with other detailed information. Further, when the destination request information simply requests the submission of information regarding the cell number of the acclimatized pluripotent stem cells as the acclimatized pluripotent stem cell information (that is, the destination simply acclimatizes to the destination). The determination unit 130 is stored in the storage unit 120 when only requesting that the cell number of the pluripotent stem cells be reported and the specific request regarding the cell number is not presented by the provider). Whether or not there is information corresponding to the destination request information based on whether or not information on the number of cells of the acclimatized pluripotent stem cells as the acclimatized pluripotent stem cell information is stored (stored) from various information. Can be determined. Further, when the provider request information simply requests the submission of information regarding the initial cell information or the acclimatization processing information (that is, the provider simply requests the provider to report the initial cell information or the acclimatization processing information). Similarly, in the case where a specific request for the initial cell information or the acclimation processing information is not presented by the provider), the determination unit 130 also selects from various information stored in the storage unit 120. , The presence or absence of information corresponding to the provider request information can be determined based on whether the initial cell information or the acclimatization processing information is stored (stored).

The determination result of the determination unit 130 regarding whether or not there is a difference between the provision destination request information and the acclimatized pluripotent stem cell information can be transmitted to the display unit 140. As a result, the administrator of the acclimatized cell management device 100 can easily confirm whether or not the acclimatized pluripotent stem cells requested by the provider can be arranged based on the determination result. That is, the administrator of the acclimatization cell management device 100 can manage whether or not the acclimatization process is appropriately executed according to the request of the provider.

As described above, the display unit 140 can display the determination result by the determination unit 130. A general monitor screen can be used as the display unit 140, and the display unit 140 is arranged in a place where the administrator of the acclimation cell management device 100 can see. For example, when the acclimatized cell management device 100 is regarded as a server device, the acclimatized cell management device 100 does not necessarily have to be provided with the display unit 140, and another terminal device owned by the administrator (for example, a smartphone). Etc.), the determination result by the determination unit 130 may be displayed on the display screen of another terminal device owned by the administrator by making it possible to connect to the acclimation cell management device 100 via the communication line. ..

The first user interface unit 150 can input various settings of the acclimatized cell management device 100 via the operation of the administrator. For example, the first user interface unit 150 relates to the above-mentioned difference between X and Y when the determination unit 130 determines whether or not the provision destination request information and the acclimatized pluripotent stem cell information are different from each other. The "determined range", the setting of detailed information (information such as the number of cells) to be focused on when the determination unit 130 executes the determination, and the like can be input via the operation of the administrator. Further, the first user interface unit 150 can input journalizing rules (classification rules) and the like of various information stored in the storage unit 120 via the operation of the administrator. Furthermore, the first user interface unit 150 can input information other than the information received from the acclimatization processing device 200 via the first communication unit 110 via the operation of the administrator.

Here, the information other than the information received from the acclimatization processing device 200 via the first communication unit 110 and input via the first user interface unit 150 includes, for example, at least the following information. One can be included without limitation. Each piece of information exemplified below is input to the acclimation processing device 200 via the second user interface unit 240 described later, and then the acclimation cell management is performed from the acclimation processing device 200 via the first communication unit 110. It may be configured to be transmitted to the device 100.
(1) Regarding the type of the target differentiated cell in the case of performing the differentiation induction treatment for inducing the differentiation of the acclimated pluripotent stem cell into a predetermined cell after the acclimation treatment with respect to the conditioned pluripotent stem cell obtained by the acclimation treatment. Information and optional information such as genetic testing as needed.
(2) Information on safety such as impurity test, sterility / mycoplasma denial test, endotoxin test, etc. when performing safety confirmation processing to carry out the specified safety test on the obtained cells.
(3) Shipping management information such as product status and transport information when the designated cells (acclimated pluripotent stem cells) are transferred to the destination.

2-2. Habituation processing device 200
As described above, the acclimation processing device 200 is connected to the acclimation cell management device 100 via a communication line, and can acquire various information and transmit the acquired various information to the acclimation cell management device 100 in real time. it can.

As shown in FIG. 3, the acclimation processing device 200 can mainly include a acclimation processing execution unit 210, a sensor unit 220, a second communication unit 230, and a second user interface unit 240.

The acclimation process execution unit 210 can include all hardware that executes the primary acclimation process and the secondary acclimation process. Specifically, the acclimation process execution unit 210 includes a culture vessel used for executing the primary acclimation process, a culture container for suspension culture for executing the secondary acclimation process, a stirring member, a culture device, a medium supply device, and the like. Can be included without limitation.

The sensor unit 220 is provided in the acclimation processing device 200 as hardware and can acquire various information. The sensor unit 220 can include various sensors such as a temperature sensor and an image sensor, for example. The sensor unit 220 changes the initial cell information about the flat-cultured pluripotent stem cells (pluripotent stem cells before being subjected to the acclimatization treatment) received from the supplier, and the pluripotent stem cells from the flat culture to the suspension culture. It is possible to obtain one or more pieces of information such as acclimation processing information regarding acclimation processing and acclimated pluripotent stem cell information regarding acclimated pluripotent stem cells generated by the acclimation process as an example. ..

Here, the initial cell information may include information on the cell origin (donor) and cell type (bone marrow, cord blood, skin fibroblast, fat, etc.) of pluripotent stem cells before being subjected to acclimation treatment. it can. As an example, such initial cell information can be adopted as a configuration in which a barcode or an individual chip is input in association with an identification ID in a culture vessel containing pluripotent stem cells before being subjected to acclimation treatment. .. As a result, when the culture vessel is placed at a predetermined position in the acclimatization processing device 200, the sensor unit 220 reads out the initial cell information input to the barcode, the individual chip, etc., so that the sensor unit 220 is initially set. Cell information can be acquired.

The acclimation processing information can include primary acclimation processing information regarding the primary acclimation processing and secondary acclimation processing information regarding the secondary acclimation processing. The primary acclimation processing information includes information on the medium used for the primary acclimation treatment and information on the culture conditions (culture container volume, seeding density, temperature, pH, number of passages, passage conditions, etc.) related to the primary acclimation treatment. sell. The secondary acclimation treatment information may include information on the medium used in the secondary acclimation treatment and information on the culture conditions (volume, temperature, pH, passage number, stirring speed, etc. of the culture vessel) regarding the secondary acclimation treatment. The acclimatization processing information may be acquired by a sensor unit 220 such as a pH sensor, and for example, information such as the volume of the culture vessel and the number of passages is separately input by the administrator via the second user interface unit 240. You can do it.

The acclimated pluripotent stem cell information includes primary conditioned pluripotent stem cell information on primary conditioned pluripotent stem cells and secondary conditioned pluripotent stem cell information on secondary conditioned pluripotent stem cells. Can be included. The primary conditioned pluripotent stem cell information may include information on the number of cells of the primary conditioned pluripotent stem cells, cell morphological characteristics, and the like. The secondary conditioned pluripotent stem cell information may include information on the number of cells of the secondary conditioned pluripotent stem cells, cell morphological characteristics, and the like. In addition, the primary conditioned pluripotent stem cell information may include biochemical information relating to the primary conditioned pluripotent stem cells. Similarly, the secondary conditioned pluripotent stem cell information may include biochemical information relating to the secondary conditioned pluripotent stem cells.

Here, the number of cells of the pluripotent stem cell that has undergone the primary acclimation treatment and the number of cells of the pluripotent stem cell that has undergone the secondary acclimation treatment can be obtained by the sensor unit 220 as an image sensor, or by another method. It can also be obtained based on. Similarly, the characteristics of the cell morphology can also be acquired by the sensor unit 220 as an image sensor. The information regarding the characteristics of the cell morphology can include, for example, information such as the number and size of the cell mass, the shape of the cell mass, and the shape of the cell surface.

Since the sensor unit 220 can monitor the acclimation processing device 200 in real time, various information exemplarily listed as described above can be obtained in real time. The sensor unit 220 can transmit all the obtained various information to the second communication unit 230.

Further, the sensor unit 220 can also acquire the status of the acclimatization process executed by the acclimatization process execution unit 210 as image information or moving image information.

The second communication unit 230 can transmit various information received from the sensor unit 220 to the acclimation cell management device 100 (the first communication unit 110) in real time. As a result, the administrator can manage in real time how the acclimation process is executed via the display unit 140 of the acclimation cell management device 100. More specifically, the administrator manages and monitors various information in real time via the display unit 140 of the acclimatization cell management device 100, so that even if some problem occurs in the acclimatization process. , You can take immediate action.

The second user interface unit 240 can input various settings related to the acclimatization process executed by the acclimatization process execution unit 210 via the operation of the administrator. Further, the second user interface unit 240 can acquire the above-mentioned initial cell information, acclimatization processing information, acclimatized pluripotent stem cell information and the like (for example, primary acclimatization that can be acquired based on the fluorescent staining method) that cannot be acquired by the sensor unit 220. The number of treated pluripotent stem cells and the number of cells of the secondary acclimation-treated pluripotent stem cells) can be input via the operation of the administrator.

As described above, the acclimatized cell management device 100 and the management system 1 including the acclimatized cell management device 100 according to the embodiment include information on the pluripotent stem cells subjected to the acclimatization process, information on the acclimatized pluripotent stem cells, and the acclimatization process itself. By acquiring various information such as information, the habituation process can be appropriately managed and monitored. This makes it possible to provide high-quality and highly reproducible acclimatized pluripotent stem cells to the recipient (customer). Furthermore, by not only providing acclimatized pluripotent stem cells to the recipients, but also providing various information associated with them, we provide additional value to the recipients. It becomes possible to do.

Further, in the above-mentioned management system 1, the acclimation cell management device 100 and the acclimation processing device 200 do not necessarily have to be physically arranged in the same place (site). That is, it is also possible to manage and monitor the acclimation process of the acclimation processing device 200 by the remote acclimation cell management device 100. Therefore, for example, the acclimatization processing device 200 is lent to the provider, and the actual acclimatization process is executed by the provider, while the administrator of the acclimatization processing device 200 remotely manages, monitors (or needs) the acclimatization process. It will also be possible to provide support).

This application is based on the following Japanese patent application and enjoys the priority of the Japanese patent application. In addition, the entire contents of the following Japanese patent application are incorporated herein by reference.
(1) Japanese Patent Application No. 2019-150483 submitted on August 20, 2019, entitled "Method for Producing Acclimated Cells"

All documents, patent applications, and technical standards described herein are incorporated herein by reference in their entirety.

Claims (29)

1. Receiving flat-cultured pluripotent stem cells from a supplier,
   Perform acclimatization treatment on the received pluripotent stem cells to enable the transition from planar culture to suspension culture.
   By the acclimation treatment, pluripotent stem cells acclimated to suspension culture are obtained.
   To provide the obtained conditioned pluripotent stem cells to a designated destination in response to a request from the destination.
   A method for producing acclimated cells, including.
2. The acclimation process
   The received pluripotent stem cells are subjected to primary acclimation culture in a primary acclimation culture vessel filled with a primary acclimation medium to obtain primary acclimated pluripotent stem cells.
   The primary conditioned pluripotent stem cells are subjected to suspension culture in a suspension culture container filled with a secondary acclimation medium to obtain secondary conditioned pluripotent stem cells acclimated to the suspension culture.
   The method for producing an acclimated cell according to claim 1, which comprises.
3. Regarding initial cell information about the pluripotent stem cells before being subjected to the acclimation treatment, primary acclimation processing information regarding the primary acclimation treatment, secondary acclimation processing information regarding the secondary acclimation treatment, and the primary acclimated pluripotent stem cells. To obtain at least one information selected from the group consisting of primary conditioned pluripotent stem cell information and secondary habituated pluripotent stem cell information relating to the secondary conditioned pluripotent stem cell.
   The method for producing conditioned cells according to claim 2, further comprising.
4. The method for producing an acclimated cell according to claim 3, wherein the initial cell information includes at least information on the cell type of the pluripotent stem cell before being subjected to the acclimation treatment.
5. The acclimated cell preparation method according to claim 3, wherein the primary acclimation treatment information includes at least information on the primary acclimation medium and information on culture conditions related to the primary acclimation treatment.
6. The acclimated cell preparation method according to claim 3, wherein the secondary acclimation treatment information includes at least information on the secondary acclimation medium and information on suspension culture conditions related to the secondary acclimation treatment.
7. The primary conditioned pluripotent stem cell information includes at least information on the cell number and cell morphological characteristics of the primary conditioned pluripotent stem cells.
   The method for producing an acclimated cell according to claim 3, wherein the secondary acclimated pluripotent stem cell information includes at least information on the number of cells of the secondary acclimated pluripotent stem cell and the characteristics of cell morphology.
8. At least one acquisition selected from the group consisting of the initial cell information, the primary acclimation processing information, the secondary acclimation processing information, the primary acclimated pluripotent stem cell information, and the secondary acclimation pluripotent stem cell information. To provide the provided information to the above-mentioned destination,
   The method for producing an conditioned cell according to any one of claims 3 to 7, further comprising.
9. The method for producing an acclimated cell according to any one of claims 1 to 8, wherein the acclimated pluripotent stem cell is provided to the donor in a suspension culture form.
10. A method of acclimatization cell management performed by at least one processor that executes computer-readable instructions.
    When the at least one processor executes the instruction,
    Acquire and store early cell information about planar cultured pluripotent stem cells,
    For the pluripotent stem cells, the acclimatization process information regarding the acclimatization process for enabling the transition from the planar culture to the suspension culture is acquired and stored.
    A method for managing acclimated cells, which acquires and stores acclimated pluripotent stem cell information regarding the acclimated pluripotent stem cells generated by the acclimation process.
11. By executing the instruction, the at least one processor further
    The destination request information regarding the request for the conditioned pluripotent stem cell of the destination to which the conditioned pluripotent stem cell is provided is acquired and stored in advance.
    The conditioned cell management method according to claim 10, wherein the provided destination request information is compared with the acclimated pluripotent stem cell information to determine whether or not there is a difference between the two.
12. By executing the instruction, the at least one processor further
    Provided by which the acclimated pluripotent stem cell is provided with at least one information selected from the group consisting of the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information via a communication line. The conditioned cell management method according to claim 10 or 11, which is transmitted to the destination server device.
13. Any of claims 10 to 12, wherein the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information are acquired from the acclimation processing device that executes the acclimation processing via a communication line. The conditioned cell management method according to paragraph 1.
14. The acclimatization cell management method according to claim 11, wherein the provision destination request information is acquired from the provision destination server device of the provision destination via a communication line.
15. The acclimation process
    The pluripotent stem cells are subjected to primary acclimation culture in a primary acclimation culture vessel filled with a primary acclimation medium to obtain primary acclimated pluripotent stem cells.
    The primary conditioned pluripotent stem cells are subjected to suspension culture in a suspension culture container filled with a secondary acclimation medium, and a secondary acclimation treatment for obtaining secondary conditioned pluripotent stem cells acclimated to the suspension culture is performed. Including
    The acclimation processing information includes primary acclimation processing information regarding the primary acclimation processing and secondary acclimation processing information regarding the secondary acclimation processing.
    The conditioned pluripotent stem cell information includes the primary conditioned pluripotent stem cell information regarding the primary conditioned pluripotent stem cell and the secondary conditioned pluripotent stem cell information regarding the secondary conditioned pluripotent stem cell. The conditioned cell management method according to any one of claims 10 to 14, which comprises.
16. The conditioned cell management method according to any one of claims 10 to 15, wherein the initial cell information includes at least information on the cell type of the pluripotent stem cell.
17. The conditioned cell management method according to claim 15, wherein the primary acclimation treatment information includes at least information on the primary acclimation medium and information on culture conditions related to the primary acclimation treatment.
18. The acclimation cell management method according to claim 15, wherein the secondary acclimation treatment information includes at least information on the secondary acclimation medium and information on suspension culture information on the secondary acclimation treatment.
19. The primary conditioned pluripotent stem cell information includes at least information on the cell number and cell morphological characteristics of the primary conditioned pluripotent stem cells.
    The conditioned cell management method according to claim 15, wherein the secondary conditioned pluripotent stem cell information includes at least information on the cell number and cell morphological characteristics of the secondary conditioned pluripotent stem cells.
20. Equipped with one or more processors
    The processor
    Acquire and store early cell information about planar cultured pluripotent stem cells,
    Acquire and store the acclimation process information regarding the acclimation process for performing the acclimation process to enable the transition from the planar culture to the suspension culture for the pluripotent stem cells.
    Acquires and stores the acclimated pluripotent stem cell information regarding the acclimated pluripotent stem cell generated by the acclimation process.
    An acclimatized cell management device that causes the at least one processor to function as described above.
21. The donor request information regarding the request for the conditioned pluripotent stem cell of the donor to which the conditioned pluripotent stem cell is provided is acquired and stored in advance.
    The destination request information is compared with the acclimatized pluripotent stem cell information to determine whether or not there is a difference between the two.
    The conditioned cell management device according to claim 20, wherein the at least one processor is further functioned as described above.
22. Provided that the acclimated pluripotent stem cell is provided with at least one information selected from the group consisting of the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information via a communication line. Send to the destination server device,
    The acclimated cell management device according to claim 20 or 21, wherein the at least one processor further functions as described above.
23. Any of claims 20 to 22, wherein the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information are acquired from the acclimation processing device that executes the acclimation processing via a communication line. The acclimated cell management device according to paragraph 1.
24. The acclimatized cell management device according to claim 21, wherein the provider request information is acquired from the provider server device of the provider via a communication line.
25. By running on at least one processor
    Acquire and store early cell information about planar cultured pluripotent stem cells,
    Acquire and store the acclimation process information regarding the acclimation process for performing the acclimation process to enable the transition from the planar culture to the suspension culture for the pluripotent stem cells.
    Acquires and stores the acclimated pluripotent stem cell information regarding the acclimated pluripotent stem cell generated by the acclimation process.
    A computer program that activates at least one processor as described above.
26. The donor request information regarding the request for the conditioned pluripotent stem cell of the donor to which the conditioned pluripotent stem cell is provided is acquired and stored in advance.
    The destination request information is compared with the acclimatized pluripotent stem cell information to determine whether or not there is a difference between the two.
    25. The computer program of claim 25, which further causes the at least one processor to function.
27. Provided that the acclimated pluripotent stem cell is provided with at least one information selected from the group consisting of the initial cell information, the acclimation processing information, and the acclimated pluripotent stem cell information via a communication line. Send to the destination server device,
    25. The computer program according to claim 25 or 26, which further functions the at least one processor as described above.
28. Any of claims 25 to 27, wherein the initial cell information, the acclimatization processing information, and the acclimatized pluripotent stem cell information are acquired from the acclimatization processing apparatus that executes the acclimatization processing via a communication line. The computer program described in paragraph 1.
29. The computer program according to claim 26, wherein the provision destination request information is acquired from the provision destination server device of the provision destination via a communication line.

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