Method and apparatus for culturing biological cells

Abstract

The present invention relates to a method for culturing different types of biological cells, in which a carrier (2) containing one or more storage chambers (3) is housed in an incubator (1), one or more cultures (S) containing cells of a common type are stored in one of the storage chambers (3), culture parameters (t1, t2, N) are assigned to the cultures (S), the tissue coordinates (C, K) of each storage chamber (3) and the culture parameters (t1, t2, N) of the cultures (S) stored therein are stored in a data processing device, and the carrier (2) is removed from the incubator (1) at predetermined time intervals to process the cultures (S) according to their individual culture parameters (C, K). According to the present invention, the data set (S) is divided into groups (G) with correlated culture parameters. The cultures (S) are subsequently processed within the groups.

Description

[Technical Field]

[0001] The present invention relates to a method and apparatus for culturing different types of biological cells, wherein a carrier is housed in an incubator and has one or more storage chambers, one or more cultures containing cells of a common type are each stored in one of the storage chambers, culture parameters are assigned to the cultures, a data set including the tissue coordinates of each storage chamber and the culture parameters of the cultures stored in that storage chamber is stored in a data processing device, and the carrier is removed from the incubator at predetermined time intervals to process the cultures according to each culture parameter. [Background technology]

[0002] Methods and devices for culturing biological cells are described in Patent Documents 1, 2, 3, 4, 5, 6, 7, and 8. The biological cells are stored in an incubator as a culture (cell culture). Such an incubator has a housing in which a plurality of carriers are arranged. Each carrier has at least one storage chamber in which a cell culture is stored. A cell culture consists of a plurality of similar cells. Usually, microplates or microtiter plates are used as carriers, and have a size of 127.8 x 85.5 mm. 2 The incubator has a fixed layout. The storage chambers of the microplate can consist of sample wells. The temperature and / or humidity inside the incubator are maintained at predetermined values. The cells can be cells of an individual, for example, a plant, an animal, or a human. However, the cells can also belong to different individuals. The cells can be of different cell types, for example, stem cells, muscle cells, brain cells, etc. The cells are removed from the incubator at preset time intervals, in particular to perform biological treatments. Nutrients can be added to the individual cultures. However, if the culture has a cell count exceeding the maximum, the culture can be split or reduced in size. However, cells can also be removed from the culture at predetermined time intervals, particularly for biological processing. Furthermore, nutrients can be exchanged or the culture can be allocated to other carriers. To keep the cell culture alive, nutrients must be supplied at predetermined time intervals. This is done within a predefined time window. The pipetting device is used to remove nutrients from nutrient containers, e.g., bottles. This is done using the pipettes of the pipetting device. In the prior art, carriers are removed one after another from an incubator and the cultures carried by the carriers are biologically treated. In this case, each type of cell is supplied with nutrients corresponding to that type. Different pipettes are used to supply different nutrients to different storage chambers. The prior art also includes U.S. Pat. Nos. 5,699,292 and 5,999,203. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] U.S. Patent No. 9,574,174 [Patent Document 2] U.S. Patent No. 9,946,834 [Patent Document 3] U.S. Patent No. 5,106,584 [Patent Document 4] Patent No. 5854418 [Patent Document 5] Chinese Patent No. 105675864 [Patent Document 6] U.S. Patent No. 8,318,492 [Patent Document 7] U.S. Patent No. 7,546,210 [Patent Document 8] International Publication No. 2017 / 143155 [Patent Document 9] U.S. Patent No. 6,146,592 [Patent Document 10] European Patent Application Publication No. 1900806 Summary of the Invention [Problem to be solved by the invention]

[0004] It is an object of the present invention to further advance the operation of devices for culturing biological cells in a manner that is convenient to use. In particular, it is provided that the operation of multi-cell systems is optimized. [Means for solving the problem]

[0005] This object is achieved according to the invention as set forth in the claims, the dependent claims not only describing advantageous further developments of the invention as set forth in the dependent claims, but also constituting independent solutions to this object.

[0006] It is first proposed that a data set is provided in the data processing device. The data set is assigned to each culture. The data set includes the tissue coordinates of the storage chamber in which the culture is located. The tissue coordinates may include an identification mark of the carrier and an identification mark of the storage chamber on the carrier. The tissue coordinates can be used to identify the spatial position of the culture, in particular, inside an incubator. The incubator may have a plurality of regularly arranged storage locations, each of which can accommodate a carrier. Each carrier can be assigned to any storage location. Additionally, the dataset includes culture parameters for each cell culture, which can be identified via tissue coordinates. The culture parameters can include information about a time window during which nutrients must be supplied to the cell culture to keep it alive. The culture parameters can include information about the type or variety of cells in the cell culture. The culture parameters can include information about nutrient type and nutrient supply.

[0007] The present invention provides a multiplexer for sequentially processing cell cultures, particularly biologically, based on a job list. The biological processing can include supplying nutrients, removing cells, replacing the medium in which the cells are stored, or adding cells again. The multiplexer can also consist of a program in an electronic controller, which can be part of a data processing device.

[0008] The method according to the invention orders the datasets by separating them into groups in which the culture parameters are correlated, for example, exactly, regionally, or partially identical or similar.

[0009] The present invention further includes exemplary embodiments in which data sets or cultures represented by data sets are divided by time window and their nutrient type. In this case, a group contains only data sets or cultures that must be supplied with the same nutrient within a period of time. The cultures within this group are biologically treated sequentially. The culture parameters can indicate a first time point from which treatment can begin and a second time point from which treatment must end. These two time points define a culture-specific time window. Each group has a group-specific time window, and the culture or dataset contained therein can be characterized by a culture-specific time window with a length that exceeds the length of the group-specific time window. The group-specific time window is located within each culture-specific time window. Different groups are treated in separate treatment phases, one after the other. Cultures containing cells of different cell types can be treated in one treatment phase but provided with the same nutrients within the same time window. However, cells of the same type can also be treated.

[0010] An exemplary embodiment of the present invention provides that different groups of cultures are stored on one carrier. While processing a job list, this carrier is removed from the incubator at different times, and each culture in a different storage chamber on the carrier is subjected to a specific biological treatment. Multiple carriers can be housed inside the incubator in a random or functional manner. For example, a carrier removed from a first storage space by a handling machine can be placed in a second storage space different from the first storage space after a specific biological treatment of at least one culture carried by it, and the tissue coordinates of the culture assigned to the carrier are changed to correspond thereto after the carrier is placed in the second storage location.

[0011] In particular, a method for culturing biological cells includes providing a plurality of cell cultures, where each cell culture has only one cell type and / or the different cell culture types are different from each other, and where the cell types have certain culture parameters, which can be cell type specific for maintaining the culture of the cell type alive over an extended period of time through periodic biological treatments.

[0012] In one aspect of the present invention, different cell types can have matching or correlated culture parameters, allowing cultures of different cell types to be specifically biologically processed in a common processing phase. A multiplexer according to the present invention is used to order data sets by groups, which are ordered by the time point at which the biological processing begins. After the time point at which the biological processing begins, all cultures belonging to that group are biologically processed sequentially.

[0013] In an exemplary embodiment of the invention, this is done by sequentially removing all carriers with each group of cultures to be processed from the incubator. This can be done automatically using a handling machine. The cultures are processed in a processing module that is spatially separated from the incubator. For example, a pipetting device can be used to supply liquid nutrients from nutrient containers to the cultures. In a processing phase in which one group of cell cultures is processed, the nutrient containers are not changed, or only nutrient containers containing the same nutrients are changed. In a subsequent, different processing phase, a different group of cell cultures is biologically processed. This can be done by supplying different nutrients from different nutrient containers.

[0014] The present invention further includes exemplary embodiments in which a first group of cell cultures is stored in a first storage chamber of a carrier and a second group of cell cultures different from the first is stored in a second storage chamber, and the carrier is removed from and returned to an incubator to biologically treat the first group of cell cultures in a first processing phase, and removed from and returned to the incubator to biologically treat the second group of cell cultures in a second processing phase, and the carrier can then be located in different locations within the incubator.

[0015] The device according to the invention comprises at least one incubator and carriers housed therein, at least some of which have one or more storage chambers in which biological cells are stored. The device also comprises a data processing device, which may have a controller capable of adjusting the incubator and, in particular, the temperature or humidity in the incubator to a target value. The controller can be used to operate a handling machine that transports the carriers housed in the incubator. The device may also have a processing module, in which biological processing of the cell cultures transported by the carriers can be performed.

[0016] According to the invention, each cell culture has a data set assigned to it, which data set is stored in the data processing device, for example as a table. The data set includes tissue coordinates used to identify the storage location of the cell culture. The tissue coordinates may include information about where the carrier is housed and the location of the storage chamber in which the cells are placed. The tissue coordinates serve to identify the associated culture with respect to its location. The data set further includes culture parameters including information necessary for the biological treatment. These may include information about nutrient type and nutrient supply. The information may indicate the earliest possible time at which the treatment must be started and the latest time at which the treatment must be completed. In addition to qualitative characteristics of the nutrients, the information may also include quantitative information about the nutrients.

[0017] Culture parameters of data sets or cultures assigned to a common group may be correlated with one another, e.g., coincident. Nutrient-related culture parameters are particularly coincident. However, correlation of culture parameters may also be understood to mean that the culture parameters have a common overlap, e.g., represent a time window bounded by two time points. However, correlation of culture parameters may also be understood to mean that the culture parameters include range specifications, and a common range exists within the range of all data sets. Two culture parameters are said to be correlated with one another if a common time window may exist within the individual time windows of the two culture parameters. Another culture parameter can be the duration of the period between two treatments. After one treatment, the range specification, which indicates the absolute time range, can be updated by adding the length of the period to the time point of the last treatment, and the time point thus obtained can define the position of the time window, e.g., the midpoint of the time window.

[0018] According to the invention, a table is arranged to be sorted by culture parameters, the table containing groups of correlated data sets, e.g., similar, regionally consistent, or having the same culture parameters, and these groups are ordered relative to each other by the time at which the biological treatment must be initiated.

[0019] The culture parameters may not only consist of information about the time point at which the biological treatment is started, i.e., the time point at which the nutrient supply is started. They may also include information about the maximum length of the time interval during which the nutrient supply must be carried out. Within a group, there may be datasets with different time intervals but with the same value for the start time of the nutrient supply. The datasets are sorted within a group so that the dataset with the shortest time interval is processed at the beginning of the treatment phase. As a result, the datasets can be sorted within a group by time interval. In particular, each culture parameter has a tuple of numerical values. Preferably, the carrier comprises a microplate as described above, which has a plurality of sample wells arranged in a grid pattern, each sample well being a storage chamber for holding a culture.

[0020] An advantage of the method or the device according to the invention is that carriers carrying different cell cultures but with correlated culture parameters can be removed one after the other from the incubator during one treatment phase and transported to a treatment module for biological treatment of the cell cultures in the same way, after which the carriers are returned to the incubator, and the nutrients contained in the treatment module are not changed during that treatment phase. As a result, the method according to the invention allows for economical biological treatment of cultures in a multi-culture system. [Brief explanation of the drawings]

[0021] The invention is explained in more detail below on the basis of exemplary embodiments. [Figure 1]FIG. 1 is a schematic table of 12 data sets of 12 cultures S stored in 6 individual chambers of two carriers C, with information on the culture parameters: the earliest time t1 at which treatment can begin, the latest time t2 at which treatment must end, and nutrients N. [Figure 2] FIG. 2 is a table showing a timetable 9 having four groups I, II, III, IV that are processed sequentially. [Figure 3] FIG. 3 is a table showing a job list 10 on the basis of which the group G of cultures 2 shown in FIG. 2 is processed sequentially. [Figure 4] FIG. 4 is a schematic diagram of a processing device comprising an incubator 1, a carrier 2 with a storage chamber 3 arranged inside, and a handling machine 5 that transports the carrier 2 to a processing module 6, where the culture arranged in the storage chamber 3 is supplied with nutrients contained in bottles using a pipetting device 7. DETAILED DESCRIPTION OF THE INVENTION

[0022] FIG. 4 shows a schematic diagram of an apparatus with an incubator 1, which includes a closed container and can be set to a predetermined temperature and / or humidity. The incubator 1 has multiple storage compartments that can be fitted with identically designed carriers 2. In the present invention, the carriers 2 are shaped like microplates. Each carrier 2 has multiple storage chambers 3, preferably sample wells. To keep the cells of the cell culture stored in the storage chambers 3 alive for a long period of time, the carriers 2 are removed from the incubator 1 at predetermined time intervals. This is done by a handling machine 5. The carriers 2 are transported to a processing module 6, which is equipped with a pipetting device 7 with one or more pipettes. The one or more pipettes are used to remove liquid nutrients N from bottles 8 and dispense them into the storage chambers 3. Reference numeral 4 denotes a data processing unit, which may also include a controller.

[0023] According to the present invention, a plurality of culture The objects are processed in groups. From the total number of cultures, those that must be supplied with the same nutrient N for a short period of time are combined into a first group. For this purpose, a data set is assigned to each culture. In FIG. 1, these cultures S are numbered 1 to 12. In the exemplary embodiment, two carriers C are provided, each with six storage chambers K. As a result, each data set has two tissue coordinates C, K, which allow the culture S to be assigned to one location.

[0024] The table shown in Figure 1 further includes culture parameters t1 and t2, which indicate when a treatment can and should be performed. In addition to these culture parameters, it also includes information about the nutrient N used, of which two nutrients A and B are provided in the exemplary embodiment. Based on the culture parameters, the culture S can be assigned to four different groups G. For simplicity, only two culture parameters are shown here. Further culture parameters can also be provided, such as the type of cell, the duration between two treatments, etc.

[0025] Figure 2 shows four groups I, II, III, and IV. Each group G has a different group-specific time window ΔT during which treatment of culture S can be carried out. Furthermore, each group G has a group-specific nutrient N. Figure 2 shows a timetable 9 consisting of four consecutive treatment phases.

[0026] All cultures S assigned to one of the four groups G are characterized by their group-specific time windows lying within the respective culture-specific time windows t1, t2.

[0027] This creates the job list 10 shown in Fig. 3. According to this, groups I, II, III, and IV are processed sequentially, specifically at their respective starting points at the start time t0.

[0028] The above descriptions are intended to describe the inventions to which this application is directed as a whole, and each also independently advances the prior art by at least one combination of the following features: do.

[0029] A method characterized in that a plurality of data sets S are divided into a plurality of groups G having correlated culture parameters t1, t2, and N.

[0030] A method characterized in that a plurality of cultures S are treated in groups.

[0031] Culture parameters t1, t2, N comprises at least information about the type of nutrient N, information about the time window t1, t2 of the treatment, information about the amount of nutrient N, information about the duration between two treatments, information about the treatment type, and / or information about the cell type, and / or the treatment comprises the supply of nutrients, and / or the treatment comprises the removal of cells from the storage chamber 3.

[0032] A method characterized in that cultures S of different groups G are stored on a carrier 2.

[0033] A method characterized in that an electronic data processing device (4) creates a job list (10) containing cultures (S) to be processed in chronological order.

[0034] Method characterized in that the tissue coordinates C, K allow unambiguous identification of the carrier 2 and the storage chamber 3.

[0035] A method characterized in that, during a processing phase in which only each culture S of one group G is processed, a handling machine 5 is used according to a job list 10 to sequentially remove from an incubator 1 only carriers 2 that store at least one culture S that is located within a group-specific time window ΔT in that time window t1, t2 and that is processed in the same way and / or that store at least one culture S that is supplied with the same nutrients N within a common time window ΔT.

[0036] The apparatus is characterized in that the data processing device 4 is programmed so that the data set is divided into groups G having correlated culture parameters, and in each processing phase in which only each culture S of one group G is processed, only carriers 2 storing at least one culture S that is biologically processed in the same way and / or at least one culture S that is supplied with the same nutrients N are sequentially removed from the incubator 1 within the time window ΔT of this group.

[0037] 1. A device or method characterized in that the carrier 2 is a microplate and the storage chamber 3 is a sample well of the microplate, and / or the carrier 2 has a rectangular base with a width of 127.8 mm and a depth of 85.5 mm, and / or during the processing phase only common nutrients N are supplied to the culture S from a nutrient container 8 using a pipetting device 7, and / or a pipetting device 7 controlled by a program of a controller is used, and / or the carrier 2 is placed in the incubator 1 in a random or functional manner, and / or the culture parameters include information about the earliest processing time t1 and the latest processing time t2 for the cell type and / or the maximum number of cells in the culture S.

[0038] All disclosed features are essential to the invention (both by themselves and in combination with one another). The disclosure of the present application incorporates in its entirety the disclosures of the relevant / additional priority documents (copies of earlier applications), with the aim of incorporating the features of these documents into the claims of the present application. The dependent claims are characterized by independent, inventive further developments of the prior art, even without the features of the claims cited therein, in particular for the purpose of filing a divisional application based on these claims. The invention specified in each claim may additionally have one or more features specified in the preceding description, particularly those designated by reference signs and / or specified in the sign explanations. The present invention also relates in particular to design forms in which individual features set forth in the preceding description are not implemented, insofar as they are clearly unnecessary for the respective intended use or can be replaced by other means having the same technical effect. [Explanation of symbols]

[0039] 1. Incubator 2. Career 3 Storage Chamber 4 Data Processing Device 5 Handling Machine 6 Processing Module 7 Pipetting Device 8 bottles 9. Schedule 10 Job List t0 Start time t1 time window, processing time t2 time window, processing time A. Nutrients B. Nutrients C Carrier G Group K storage chamber N Nutrients S dataset ΔT time window

Claims

1. 1. A method for culturing different types of biological cells, comprising: A carrier (2) having one or more storage chambers (3) is housed within the incubator (1); One or more cultures (S), each having a common type of cell, are stored in one of the storage chambers (3); The culture (S) is assigned culture parameters (t1, t2, N), the culture parameters (t1, t2, N) including information about a time window (t1, t2) of a process of supplying the culture (S) with nutrients (N) to keep the culture (S) alive, information about the type of cells in the culture (S), and information about the type of nutrients (N) and the supply of the nutrients (N); a data set (S) comprising the tissue coordinates (C, K) of each storage chamber (3) and the culture parameters (t1, t2, N) of the culture (S) stored therein is stored in a data processing device (4), the tissue coordinates (C, K) comprising an identifier of the carrier (2) and an identifier of the storage chamber (3) and can be used to identify the spatial position of the culture (S) within the incubator (1); a job list (10) containing the data set (S) of the culture (S) to be processed in chronological order is created by the data processing device (4), and the carrier (2) is removed from the incubator (1) at predetermined time intervals to process the culture (S) according to each of the culture parameters (t1, t2, N) based on the job list (10); The data sets (S) are divided into groups having matching or correlated culture parameters (t1, t2, N) with respect to the time windows (t1, t2) and the type of nutrients (N); The cultures (S) having the identical or correlated culture parameters (t1, t2, N) and different cell types are biologically treated in a common treatment phase of one group, and during the treatment based on the job list (10), all cultures (S) belonging to the same group are biologically treated sequentially; The biological treatment may include supplying the culture (S) with nutrients (N) to keep the culture (S) alive, removing cells from the storage chamber (3), replacing the medium in which the cells are stored, or re-adding cells.

2. 2. The method according to claim 1, characterized in that the culture parameters (t1, t2, N) further comprise information on the amount of the nutrient (N), information on the duration between two treatments, and information on the treatment type.

3. 3. The method according to claim 1 or 2, characterized in that the cultures (S) of different groups (G) are stored on one carrier (2).

4. 4. The method according to claim 1, wherein the time window (t1, t2) in the culture parameters (t1, t2, N) of each culture (S) of one group (G) includes a group-specific time window (ΔT) for that group (G), and in one processing phase in which only each culture (S) of one group (G) is processed, only carriers (2) storing at least one culture (S) of that group (G) are sequentially removed from the incubator (1) according to the job list (10) using a handling machine (5).

5. 4. The method according to claim 1, wherein the type of nutrients (N) in the culture parameters (t1, t2, N) of each culture (S) of one group (G) is the same, and in one processing phase in which only each culture (S) of one group (G) is processed, only carriers (2) storing at least one culture (S) of that group (G) are sequentially removed from the incubator (1) according to the job list (10) using a handling machine (5).

6. An apparatus for carrying out the method according to any one of claims 1 to 5, comprising an incubator (1) and carriers (2) accommodated therein, at least one of said carriers (2) having one or more storage chambers (3) for storing biological cells, a data set (S) comprising the tissue coordinates (C, K) of each storage chamber (3) and the culture parameters (t1, t2, N) of the cultures (S) stored therein is stored in a data processing device (4); said device being provided with means for removing said carrier (2) from said incubator (1) at predetermined time intervals and treating said cultures according to respective culture parameters (t1, t2, N), Apparatus, characterized in that the data processing device (4) is programmed to carry out the method according to any one of claims 1 to 5.

7. 7. The device according to claim 6, wherein the carrier (2) is a microplate and the storage chamber (3) is a sample well of the microplate.

8. 8. Device according to claim 6 or 7, characterized in that the carrier (2) has a rectangular base with a width of 127.8 mm and a depth of 85.5 mm.

9. 9. The device according to any one of claims 6 to 8, characterized in that in one processing phase only one common nutrient (N) is supplied to the culture (S) from a nutrient container (8) by means of a pipetting device (7).

10. 10. The device according to any one of claims 6 to 9, characterized in that a pipetting device (7) is used which is controlled by a program of the control device.

11. 11. Apparatus according to any of claims 6 to 10, characterized in that the carriers (2) are placed randomly or functionally in the incubator (1).

12. 12. Apparatus according to any one of claims 6 to 11, characterized in that the culture parameters (t1, t2, N) further comprise information regarding the maximum number of cells in the culture (S).

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