CN117098836A - System for culturing biological cell cultures - Google Patents

Abstract

The invention relates to a system for culturing biological cells in a processing module (3), the biological cells being stored in a storage module (2) on a carrier (1), wherein each cell line is provided with an application (A1, A2, A3) comprising a processing rule comprising one or more work flows (W1.1, W1.2, W1.3; W2.1, W2.2, W3.3; W3.1, W3.2, W3.3) which are carried out in the processing module (3) in time intervals in sequence in a start-up time (t), respectively, wherein one work flow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) comprises one work step or more work steps which need to be carried out successively in a short period of time, wherein a control device (6) controls the execution of a plurality of applications (A1, W2.2, W1.3, W2.2, W3.3.3) in succession in the same time interval in the processing module (1.1, W3.2.3) in a time sequence. In order to be able to execute the workflows in succession in a single processing module (3) in a cost-effective manner, it is proposed that each workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) is configured with an effective function (W (t)) that is dependent on a start-up time (t) and that the order and start-up time (t) of the workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) of the different applications (A1, A2, A3) are calculated by minimizing or maximizing the sum (Sigma) of the effective functions (W (t)).

Description

System for culturing biological cell cultures

Technical Field

The present invention relates to a system and a method for culturing a biological cell culture, the system having: a plurality of carriers, each carrying one or more cell cultures; at least one storage module in which the carrier is stored; a processing module in which the cell culture carried by the carrier is processed by means of at least one processing device; a transport device by which the carrier is transported between the storage module and the processing module; and a programmed or programmable control device which has a memory device and controls the transport device and the processing device, wherein applications which are associated with a plurality of mutually different cell cultures are stored in the memory device, wherein each application is a processing rule for processing the associated cell cultures, which processing rule contains one or more processing units, hereinafter called workflows, which are executed in each case in sequence at a start-up time in a processing module at time intervals, wherein a workflow contains one or more work steps, wherein the one or more work steps need to be executed directly in succession, wherein the control device controls the simultaneous execution of a plurality of applications and in this case starts the one or more workflows of mutually different applications in succession in time sequence in the same processing module.

Background

Cells, in particular human cells, are cultivated in a plastic vessel, for example a trough of a carrier, by adding nutrients. The attached cells are typically cultured in a monolayer that is attached to the bottom of a plastic vessel or to a well in a carrier. The cells are separated from the bottom before the whole bottom is covered with cells. For this purpose, a fusion scan is performed by means of which the coverage of the substrate, for example the bottom of the carrier, by the cells is determined. Cells separated from the bottom can be diluted and transferred to new carriers or new tanks. Prior art systems and methods for maintaining such cell cultures may have an incubator, a robot for handling the carrier carrying the cells, and a treatment device by which the nutrient medium may be added or replaced.

Patent document EP 1 598 415 A1 describes a system for culturing biological cell cultures, in which cells are stored on a carrier in a holding module. The transport device comprising the gripping elements can take the carrier out of the holding module and transport it to the processing module. Where the cell culture stored on the carrier is treated by means of a treatment device. Different types of cell cultures are treated according to different treatment rules from each other. For each cell culture, for example for each vector, there can be an application which is a treatment rule for treating the associated cell culture. Such applications may include one or more workflows. Each workflow may comprise at least one work step. However, a workflow can also have multiple work steps. The various working steps of a workflow are characterized in that they must be performed directly or sequentially in succession in a short period of time. In contrast, the multiple workflows of an application are each executed at larger time intervals between individual workflows. The carrier may be in its associated storage position in the storage module between execution of the individual workflows. The number of applications may be 100 or more and the workflow assigned to the application may be stored as a digital data set in a memory means of the control device. The programmable control means coordinate the parallel execution of the plurality of applications and control one or more workflows of mutually different applications in time sequence in succession in the same processing module.

The automation system for executing the application preferably comprises at least one or more workflows in which the nutrient medium is changed or the degree of fusion is measured, which automation system is controlled by a planning program of the control device. Such a planning program creates a time plan to execute the workflow of each individual application in succession. Furthermore, the planning procedure should be able to influence the workflow. Such a time schedule generally contains a plurality of work processes, wherein the duration of the work processes and the intervals between the work processes may vary in an unpredictable manner. Since cells are living units and their characteristics, in particular the growth rate, may change in an unpredictable manner, the working process must be adapted to the corresponding individual development of the cell culture. The growth rate of cells is generally variable and difficult to predict. The growth rate of cells may vary, for example, on a batch basis based on known factors, such as transfection, lack of serum, or cryptographic factors. Thus, the characteristics of the cell culture, in particular the growth rate, must be observed in order to adapt the time course with respect to the point in time at which the workflow starts to the state of development of the cell culture.

Furthermore, the system for culturing biological cell cultures must be able to execute the application time-efficiently and to determine the start-up time of the workflow in such a way that the workflow, which may be in the processing duration of another workflow at the optimal start-up time, is moved in time in such a way that it is executed in sequence in the processing module.

Disclosure of Invention

The object of the present invention is therefore to further develop, in an advantageous manner, a system or method for culturing a biological cell culture, in which a plurality of applications with a workflow are executed simultaneously in time. The object of the invention is, in particular, to operate the system at low cost.

The object is achieved by the invention described in the claims, wherein the dependent claims are not only advantageous developments of the invention described in claim 1, but also independent solutions to the object.

A first aspect of the invention involves avoiding a time-wise collision of workflows or workflows that may have overlapping processing times under optimal conditions. Such workflows must be moved in time such that the start time of one workflow is after the end of another workflow. According to the invention, each workflow is provided with an effective function (or function) which is dependent on the start-up time.

The effective function or curve of the effective function may have an extremum at a time when the workflow would preferably be executed. Thus, at times when a workflow would be preferably performed, the effective function is minimum or maximum or the curve of the effective function has a minimum or maximum value. Preferably the minimum of the effective function. The effective function then rises at an earlier and later time. The rise may be linear or non-linear and rises more gently in the direction of the earliest start time than in the direction of the latest start time, or conversely, in the direction of the latest start time than in the direction of the earliest start time. If the workflow is, for example, a workflow in which the state of the cell culture should be monitored, the workflow may have a work step in which the coverage of the bottom of the cell to the tank storing the cells is measured. Such fusion scans can be performed by means of microscopy and automated image analysis. Cells may be extracted in a subsequent step according to the coverage in order to reduce the coverage. A liquid may be added to separate the cells from the bottom. The extraction of cells may be achieved by pipetting means. At the same time, the growth rate individually associated with (or corresponding to) the cell culture can be determined by measuring the coverage by comparing the earlier measurement with the current coverage taking into account the time elapsed therebetween. The growth rate measured by such a fusion scan enables calculation of future time points at which the next monitoring of the cell culture needs to be performed in the next workflow. The system calculates a preferred start-up time. The preferred start-up time may be, for example, a time at which 80% fusion is expected to occur based on the determined growth rate. The system may calculate a latest start-up time approximately at a location where a degree of fusion of 90% to 100% is predicted based on the determined growth rate. The latest start-up time may cause overgrowth of the surface. The system may calculate the earliest start-up time, for example at a location where the calculated fusion is between 15% and 30%, preferably 20%. Using these boundary values for the earliest start-up time, the latest start-up time and the preferred start-up time, an effective function is constructed which can be normalized, for example, with a value of 0 at the preferred start-up time and a value of 1 at the earliest start-up time or the latest start-up time. This may be a function of the preferred start-up time. However, it is also possible that the effective function increases gradually or rapidly to a maximum value, in particular at the latest start-up time, when the preferred start-up time approaches the earliest or latest permissible value. If the system contains an application with at least one workflow, respectively, which is within the processing duration of the other workflow due to its preferred start-up time, the start-up times of the two workflows are changed in the variance calculation such that the sum of the two effective functions is minimized or has a minimum value. If the system contains an application where multiple workflows overlap in their execution times, a variation calculation is performed on these workflows in which the start time of the workflows is changed so that the sum of all the effective functions has a minimum. The minimum may be a local minimum. But the minimum can also be a global minimum. In particular, it is provided that at least some workflows are configured with an earliest start time before which they are not allowed to start. However, it may also be provided that at least some of the workflows have a latest start-up time after which the workflows are not allowed to start up. Furthermore, it can be provided that the earliest start-up time and the latest start-up time are directly adjacent to the preferred start-up time, so that the workflow is only allowed to start at the preferred start-up time. The effective function related in this respect may be expressed mathematically, i.e. it has one or two very steep branches, the slope of which increases rapidly as the function approaches the earliest or latest starting value. In the simplest case, the effective function may have branches that extend linearly. But the branches of the effective function may also be represented by higher order polynomials. According to a preferred embodiment of the invention, the actual start-up time of the effective functions for the workflow is chosen such that the sum of the effective functions for the workflow is minimized, since the effective functions are increased if the workflow is executed at a start-up time different from the preferred start-up time. In this way, the effective function may be regarded as a cost function, where executing the workflow too early or too late causes a penalty or cost. The goal of the system is to minimize the sum of the effective functions or cost functions on the workflow of the application, i.e. to maximize the time efficiency of the system.

The present invention also generally relates to a planning software or planner for managing a plurality of functions having one or more workflows. In alternative designs of such planners, the functionality and goals of the planners may also be reversed. The functionality may be maximized at the point in time when the work process is initiated. The goal of the system may be to maximize the sum of the effective functions on the workflow of the application in order to maximize the time efficiency of the system.

The workflow may include at least one or more of the following work steps: storage of cell cultures, counting of living or dead cells, separation of cells from the bottom of the carrier tank, extraction of cells from cell cultures to reduce coverage orThe degree of fusion, the degree of fusion scan, the replacement of the culture medium of the cell culture, the addition or discarding of nutrients of the cell culture. In particular, it can be provided that the workflows in the application are widely distributed over time, i.e., that the workflows have a large time interval from one another. For example, one workflow may last for 30 minutes. During this time, a degree of confluence scan, replacement of the medium of the cell culture, or addition of nutrients may be performed. The time between two successive workflows may be hours or even days, wherein the intermediate time between successive workflows depends on the growth rate of the cell culture, which is recalculated continuously or respectively when the workflows are executed. It can thus be provided that in a workflow for monitoring the growth of cells, a fusion scan is first performed, by means of which the growth rate is determined from the fusion formed in the preceding workflow. The degree of fusion can then be reduced to, for example, 10% by extracting the cells. The degree of fusion can then be reduced to, for example, 10% by removing, isolating, diluting or seeding the cells into new vectors. From the determined growth rate, a preferred start-up time (t _p ) So that the predicted degree of fusion reaches 80%. Based on the determined growth rate, the effective function is also redetermined, e.g. by determining the earliest and latest start-up times (t _i ) The effective function was re-determined with 20% fusion at the earliest start-up time and 95% fusion predicted at the latest start-up time.

The work steps within the workflow form a time-dependent process which has to be performed sequentially, for example at time intervals of ideally 0 (zero) or at most a few minutes, wherein no further workflow can be inserted between the work steps. For example, the first working step may be to remove the carrier from the holding module and transport it to the processing module. The second subsequent working step may for example be a fusion scan, in which the coverage of cells attached to the bottom of the vessel is determined by optical means. The third working step may be the addition of a "separating agent" by which the cells are separated from the bottom of the vessel or the bottom of the carrier tank. The fourth working step is to at least partially remove the cells from the nutrient solution in the vessel or tank or to transfer a portion of the cells to a new vessel or tank. By this step the coverage, i.e. the degree of fusion, can be adjusted. The fifth working step may be the addition of a nutrient solution. The sixth working step may be to transport the carrier back to the save module. Each of these work steps may affect the progress of the function. Other work steps may include the counting of living and dead cells. This working step may be performed before or after the second working step. For culturing a specific cell line, such as CHO cells (chinese hamster ovary), examples of applications are created that contain parameters for culturing the cell line, such as complete media formulation, degree of fusion when cells need to pass, split ratio and number of carrier plates (petri dishes). One application may involve one or more carrier plates. For applications involving multiple carrier plates, each having multiple vessels, tanks or carriers, in which cell cultures are stored, there are workflows that last longer than applications involving only one carrier plate. The interval between the two workflows depends essentially on the cell line and mainly on the doubling time or division ratio of the cells of the cell line. The split ratio represents the degree of dilution of the cells in each workflow. Typically, the doubling time of a cell line varies from cell line to cell line and thus also from example of application for maintenance of the cell line. There are time critical workflows that must be performed to avoid excessive cell fusion.

The application may contain workflows that vary in number and order. For example, maintenance of iPSCs (induced pluripotent stem cells) typically involves a workflow for medium replacement ("feeder cells" or "refeedcells") because iPSCs factors such as FGF2 require a short half-life. Stem cells typically need to be fed every 24 hours. The frequency of "medium exchange" can vary and depends on the growth rate or division ratio of the cells. The time point of the confluence scan also depends on the growth rate of the cells, so that the confluence scan can be performed, for example, before or after the first or second medium change.

The above description characterizes the effective function as a function that needs to be minimized, for example, in the case where the effective function represents cost. However, the opposite is also possible, for example, in the case where the effective function represents a benefit. In this case, the effective functions respectively need to be maximized.

Drawings

Embodiments of the present invention are described below with reference to the accompanying drawings. In the drawings:

fig. 1 shows three applications Al, A2, A3 in the upper region indicated by a, with associated workflows w1.l, W1.2, W1.3, respectively; w2.1, W2.2, W2.3 and W3.1, W3.2 and W3.3, which need to be performed at a preferred start-up time, wherein the workflow W2.1, W2.2 and W2.3 of application A2 has to be performed at a fixed (non-modifiable) start-up time,

In the lower region, denoted by b, the workflow of the three applications is arranged after it has been partially moved in time (see arrow), so that said workflow can be executed successively in the processing module,

figure 2 shows the effective function W (t) in the upper region denoted by a,

the area below it, denoted by b, shows the time scale in days and

the lower region denoted by c shows the value of the degree of fusion corresponding to the time scale,

fig. 3 shows a method for calculating the earliest start time (t _e ) Priority start time (t _p ) And the latest start-up time (t _l ) Is used for the calculation of the graph of (a),

figure 4 shows a calculation chart also used to calculate a preferred start-up time from the fusion scan,

figure 5 shows a view according to figure 2a of the effective function W (t),

figure 6 shows a further view according to figure 2a of the effective function W (t),

fig. 7 shows an application a, which contains three cyclically repeated workflows W1, W2, W3,

fig. 8 shows an application a, which contains five workflows W1, W2, W3, W4, W5 that repeat cyclically,

FIG. 9 schematically illustrates steps for resolving a time conflict in the event that two or more workflows need to execute simultaneously or the start time of one workflow is within the execution time of another workflow;

Figure 10 schematically shows elements of a system according to the invention,

fig. 11 schematically shows a graphical interface (user interface) for manually designing an effective function,

fig. 12 shows a view according to fig. 1, in which two workflows W2.1 and W3.2 are exchanged in their time sequence, in order to optimize the effective function, for example to maximize or minimize the effective function,

fig. 13 shows a view according to fig. 2, but for the case that the effective function illustrates the benefit and needs to be maximized.

Detailed Description

Fig. 10 schematically shows a system consisting of a preservation module 2 for preserving cell cultures. Such a preservation module 2 may be an incubator. In the housing of such a preservation module, there may be a plurality of "hotels" in which a large number of carriers 1 are stored one above the other. For such a preservation module 2 reference may be made to WO 2020/098960 Al.

A transport device 5 is provided, which transport device 5 can have grippers, end effectors and/or other transport means, for example transport rails or automation means, by means of which the carrier 1 can be transported from the holding module 2 to the processing module 3. For a design of the treatment module, reference may be made to WO 2020/098957 A1. In the processing module 3 there are one or more processing means. The handling device 4 may have a microscope, a pipetting device or the like.

The preferred plurality of carriers 1 stored in the holding module 2 may be microwell plates. Such microplates have a plurality of wells or wells in which a nutrient with a cell culture is stored, respectively. Each carrier 1 may have a cell culture of one cell type. Different vectors 1 may carry different cell lines. The holding module 2 is designed such that the cells stored therein increase at a cell-specific division rate.

The control device 6 may have a microcontroller or microcomputer which is able to control the transport device 5 and the processing device 4. The control device 6 may also have a memory device 7 in which the application is stored in digital form. The application is a treatment rule for treating cell cultures, wherein it can be provided that each carrier 1 is provided with an application, i.e. a treatment rule, which takes into account the characteristics of the cell lines carried by the carrier 1. However, an application a can also comprise a plurality of carriers 1, wherein in this case it is provided in particular that a plurality of carriers 1 carry cells of the same type. In the storage module 2, carriers 1 can be stored which each carry a cell culture system that differs from one another and are each provided with a different application Al, A2, A3. Each application may have one or more workflows W1, W2, W3, W4, W5, where each workflow may contain one or more work steps. The workflows applying Al, A2, A3 need to be executed in sequence at preset time intervals, wherein each workflow W1, W2, W3 has a preferred start-up time at which the workflow can be started. However, some or all of the workflows W1, W2, W3 may also be started at an earlier or later point in time, but not at the earliest start time t _e Before and not at the latest start-up time t _l And then started. There may be times between minutes and days between the workflows of the application. There is preferably a time between hours to six days, half day to six days, or between one and four days between the workflows. The duration of the workflow may be between one minute and 60 minutes.

A workflow comprises one or more work steps that must be performed directly or sequentially in a short time, wherein two sequentially sequential work stepsThe length of the pause between the work steps may preferably be zero or typically a few minutes, during which there are no other workflows. One working step may be, for example, a fusion degree scan 11 in which the measured fusion degree 12 is determined in order to determine the preferred start time t _p . Other working steps may include adding a separating agent 14, suspending 15 the cells, centrifuging, counting 17 the living or dead cells, seeding 18 the cells, adding nutrients, i.e. feeder cells 19. The workflow may include at least one of these work steps or other work steps not previously listed.

Applications with one or more workflows W1, W2, W3, W4, W5 can be executed periodically at indeterminate times, for example for cell line maintenance. One application may last for several days.

The system may have multiple instances of the same application, for example a cell line maintenance application may have a first instance of "maintaining CHO cells" or a second instance of "maintaining Vero cells".

Fig. 7 shows an application a consisting of three workflows W1, W2, W3, wherein a first workflow W1 is executed after one to three days after the application is started, in which first workflow the fusion scan 11 is executed. After a further one to three days, a second workflow W2 is performed in which, first, a degree of fusion scan 11 is also performed and the cells are washed and a separating agent 14 is added according to the degree of fusion in order to separate the cells from the bottom of the grooves of the carrier. The fusion scan 11 lasts approximately 1 minute, while subsequent steps, such as washing or adding the separating agent 14, may take 10 to 20 minutes. The original degree of fusion may be measured in the first workflow W1. By further measuring the degree of fusion achieved in the second workflow W2, the rate of division of the cell line can be determined. From this, the preferred start time t for the next workflow can be calculated _p For example, at an expected fusion of 80%.

The third workflow W3 may also be performed after five to ten minutes. The workflow may include the steps of extracting cells 15, centrifuging 16, counting of living and dead cells 17 and/or seeding 18 of cells or seeding 18 cells 18 into new microplates 18.

Fig. 8 shows another embodiment of an application, in which five workflows W1, W2, W3, W4, W5 are executed successively, and the sequence of the five workflows repeats for an indefinite time.

Workflow W1, workflow W2 and workflow W3 may each include the addition 19 of nutrients. Workflow W6 includes the step of fusion scan 11. For example, the interval between nutrient additions may be set to 24 hours and the cell doubling time may vary from cell line to cell line, so the number of nutrient additions may vary depending on the growth rate of the cells, i.e. the number of workflows in the application. The order of the workflows W1, W2, W3, and W6 may also vary (i.e., W6 may not proceed in the order shown in FIG. 8) depending on the point in time of initiation of the fusion scan 11 and the growth rate of the cells. Workflow W4 includes a fusion scan 11 and step 14 includes removing media and adding a separating agent. The procedure 5 may comprise suspending the cells in step 15, centrifuging in step 16, counting dead and living cells in step 17 and seeding the cells in step 18.

Fig. 2 and fig. 5 and 6 show exemplary trends of the effective function W (t). The value presented by the effective function depends on the start time t of the workflow W1, W2, W3, W4, W5. The effective function W (t) is at a preferred start time t of the workflow corresponding to said effective function _p With a minimum value. The effective function corresponds to the latest start time t _l Or earliest start time t _e Has a maximum value.

Region b of fig. 2 illustratively presents the number of days corresponding to the start time given in region a. Region b of FIG. 2 shows the degree of fusion associated, i.e.the coverage of the bottom of the tank in which the cell culture is stored, as a percentage value. The optimal start-up time is the time at which the degree of fusion is expected to reach 80%. The latest start-up time is the time at which the degree of fusion is still slightly below 100%. The earliest start-up time TE is the time at which the degree of fusion is expected to be about 20% to 30%.

The control device 6 is programmed such that each workflow W1, W2, W3, W4, W5 is started at the preferred start-up time t in the absence of a conflict _p Starting. In case of a conflict where two or more workflows have to be executed simultaneously according to a time schedule preset by applying Al, A2, A3, the workflows are moved such that they can be executed sequentially in the same processing module 3. For this purpose, the workflow is moved such that the sum of the effective functions has a minimum value or is minimal. Such movement may include a change in order, i.e., rearrangement of the workflow [ ] Or working process).

Fig. 12 shows exemplarily the effective functions maximized according to the needs in the view of fig. 2.

FIG. 1 shows an example in which three applications Al, A2, A3 have three workflows W1.1, W1.2, W1.3, respectively; w2.1, W2.2, W2.3 and W3.1, W3.2, W3.3. The workflows W1.1, W2.1 and W3.1 applying Al, A2 and A3 are in such a position in time that they overlap when they are executed as planned. The workflow W2.1, W2.2 and W2.3 of the application A2 also has the feature that the workflow is only allowed to be executed at a preferred start-up time, so that the latest start-up time and the earliest start-up time are to some extent identical. These workflows W2.1, W2.2 and W2.3 control (or manage) to some extent the scheduling of the remaining workflows. Thus, a system according to the invention may comprise workflows W2.1, W2.2 and W2.3, which control the timing of the remaining workflows.

Thus, to minimize the sum of the effective functions of the workflows W1.1, W2.1 and W3.1, the workflow W1.1 moves forward in time and the workflow W3.1 moves backward in time.

In the described embodiment, the respective second workflows W2.2 and W3.2 do not need to be moved, as they can be performed sequentially as planned. Only the workflow W1.2 has to be moved slightly forward. The workflow W1.3, W2.3 and W3.3 need not be shifted in time. While the order of the workflows varies in the example shown, it is envisioned that in some cases, minimizing or maximizing the sum of the effective functions may result in a variation in the order of the workflows.

Fig. 11 shows that it may also be possible to exchange workflows W3.2 and W2.1.

FIG. 3 shows a plot of proliferation rate of cell cultures over time. Preferred start time point t of workflow _p Alternatively, the degree of fusion is thus predicted to be 80%. Last possible start time point t of workflow _l Alternatively, the degree of fusion is thus predicted to be 90%. Earliest start time point t of workflow _e Alternatively, the degree of fusion is thus predicted to be 20%.

Fig. 4 shows the calculation path of the preferred start time 13 calculated using the measured fusion 12 at the point in time of the fusion scan 11. 60% fusion has been measured at the time point of fusion scan 11. The point on the growth curve where the degree of fusion was 80% was determined.

In this example, the system counts resuspended cells and calculates the dilution, for example, when the workflow was previously performed, in order to set the degree of fusion of the cells to 10%. Based on the set 10% confluence, the system then calculates the time point at which the cells are expected to be in the middle or terminal (50% to 80% confluence) of the growth cycle. From an original degree of fusion of, for example, 10%, a time for which the degree of fusion is, for example, 80% is calculated assuming, for example, an exponentially increasing curve.

For cases where the system does not know the doubling time of the cells, the shortest known doubling time or a doubling time less than the shortest known doubling time of mammalian cells in culture may be used conservatively. For example, the system may use a temporal doubling time of 12 hours to calculate when a fusion scan should be performed. Alternatively, however, the system can also perform a fusion scan only once every 24 hours until the cells have reached a stable doubling time. Once the system has data on doubling time, the system can use the doubling time of the cells to determine when 80% confluency is reached.

Fig. 5 and 6 show variants of the effective function, wherein the effective function W (t) has a nonlinear course. In the case of the effective function shown in FIG. 5, the latest start-up time t _l And a preferred start-up time t _p The time interval between them is significantly smaller than the earliest start time t _e And a preferred start-up time t _p Time interval between.

In contrast, FIG. 6 shows the effective function W (t) relative to t _p And (5) symmetrically running.

Fig. 11 shows, as an example, a graphical interface by means of which the trend of the efficiency function W (t) can be preset manually by "dragging" the anchor point on the screen.

It may be desirable for a user to be able to add an application, workflow, or other work process to the system, which may be another more demanding planning problem that the system needs to handle. The time in the time plan may be modified. In particular, it is contemplated that some cell lines may grow faster or slower than expected. During a workflow, the processing means 4 may process one or more carrier plates directly in succession. This is preferably achieved in the same working steps.

According to the present invention, a cell culture system is provided. In one implementation, the cell culture system has an incubator, a refrigerator, two storage chambers for laboratory equipment at room temperature, and a processing module 3. The processing module 3 may have liquid handling means, such as a pipette. Other mechanisms may also be provided, such as a decapper for uncapping bottles, a gripper for handling the carrier 1, and for example a microscope. Depending on the application and workflow or other working process, the capacity of the system may be limited by the capacity of the incubator for the carrier, in particular the carrier plate, and in particular the number of plates that the incubator can handle, or by the capacity (speed) of the processing module. Thus, only a limited number of work processes or workflows are possible in the processing module 3.

The system may perform applications including, for example, cell line maintenance or cell-based "analysis" or experimentation. In terms of cell line maintenance, the application may run for an indefinite period of time, in particular may last days or weeks.

The workflow includes, inter alia, "fusion scan", "addition of separating agent" and cell extraction. The workflow of cell extraction may comprise individual operations, namely extraction of cells and seeding of cells. Furthermore, working steps such as centrifugation or counting of dead and living cells can be provided. For the culture of a specific cell line, such as CHO cells, an example of an application is created, which example contains parameters for the culture of the cell line. The parameter may be the complete medium formulation, the degree of channel fusion, the split ratio or the number of carriers, for example carrier plates such as microwell plates.

The working steps within the workflow are continuous and have no gaps between the two working steps, in particular no gaps of more than one minute. The interval between two workflows in the same application is more than one minute and typically several minutes, hours or days. If the degree of fusion required for this has not been achieved, the transport of the carrier from the holding module 2 to the processing module 3 and the processing there can be delayed. Thus, a workflow may contain work steps that are optionally performed, for example, when a fusion scan reaches a particular value. If the fusion scan does not reach a particular value, one or more of the following work steps may be omitted or deferred.

The method based on the system according to the invention is able to handle time schedules with different durations and intervals of the workflow. Referring to fig. 9, the method can be summarized as follows:

at the preferred starting point t _p Defining a workflow, defining an earliest start time t _e And a latest start-up time t _l For at a different from t _p The cost of executing the workflow, wherein the cost is represented in the effective function W (t) (executing the workflow 21).

If an input is made by the user or if data is found that would change the preferred start time of the next workflow, for example, then the data (start time) for all the next workflows is recalculated (22). The calculation is carried out such that the sum, in particular the sum, of the effective functions of the workflow reaches (23) a minimum. The minimum value is not necessarily a global minimum value, but may be a local minimum value.

The sum Σ of all the effective functions W (t) of all the workflows can be expressed as a multidimensional function.

∑＝W ₁ (t ₁ )+W ₂ (t ₂ )+…+W _n (t _n )

By shifting the respective start-up time and taking into account the duration of each workflow, a global or local minimum can thus be calculated,

the minimum value is subject to the boundary condition that all workflows succeed in time and that the start-up time point of each workflow is not within the duration of the other workflows.

The application provides a method for mixing workflows in which the sum of the effective functions W (t), which for example represents the costs that are required to execute the workflow, is minimized, and the method according to the application is therefore a cost-optimized method.

The application thus relates to a planner or planning software by means of which a plurality of applications (Anwendungen, german) can be managed, wherein each application comprises one or more workflows (work processes). The working process of the different applications is optimally performed, wherein the optimization features described before are used.

The above description serves to illustrate the application which is encompassed by the application as a whole and which also extends the prior art independently of one another at least by the following feature combinations, two, more or all of which can also be combined, namely:

a system characterized by each workflow W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3 is configured with an effective function W (t) related to the start-up time t and by minimizing the sum Σ of the effective functions W (t) the workflow W1.1, W1.2, W1.3 of the different applications A1, A2, A3 is calculated; w2.1, W2.2, W2.3; the order of W3.1, W3.2, W3.3 and the start-up time (t).

A method, characterized in that each workflow W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3 is configured with an effective function W (t) related to the start time t and by minimizing the sum Σ of the effective functions W (t) the workflow W1.1, W1.2, W1.3 of different applications A1, A2, A3 or instances of the same application is calculated; w2.1, W2.2, W2.3; the order of W3.1, W3.2, W3.3 and the start-up time (t).

A system or method, characterized by each workflow W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3 are configured with a preferred start-up time t _p In the case of the preferred start-up times, the workflow W1.1, W1.2, W1.3 is assigned; w2.1, W2.2, W2.3; the effective functions W (t) of W3.1, W3.2, W3.3 have a minimum value.

A system or method, characterized in that at least some of the first workflows W1.1, W1.2, W1.3 of the different applications A1, A2, A3 have a first preferred start-up time t _p The first preferred start time is in the duration of the processing of the second workflow, the second preferred start time t of the second workflow _p At a first preferred start-up time t _p Previously, the control device 6 has shifted the first and/or second activation times in time in such a way that the two first and second workflows W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3 are performed in sequence, wherein the first and second start-up times are selected such that the sum Σ of the effective functions W (t) is minimized or maximized.

A system or method, characterized in that at least some of the workflows W2.1, W2.2, W2.3 are configured with an earliest start-up time t _e Not allowed until the earliest start-up timeStart the workflow W2.1, W2.2, W2.3, and/or be configured with the latest start time t _l After the latest start-up time, the start-up of the workflows W2.1, W2.2, W2.3 is not allowed, wherein it is provided in particular that the effective function W (t) is at the earliest start-up time t _e And a latest start-up time t _l Where has a maximum value.

A system or method, characterized in that the effective function W (t) has a starting time t from earliest _e Linear or non-linear to the preferred start-up time t _p Descending or ascending branch and slave preferred start-up time t _p To the latest start time t _l Branches that rise or fall linearly or nonlinearly.

A system or method, characterized in that the trend of the effective function W (t) is selected such that the costs that are spent in the workflows W1.1, W1.2, W1.3 for executing the applications A1, A2, A3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3 at the preferred start-up time t _p There is a minimum in the case of start-up and a maximum in the case of start-up of the workflow at the earliest start-up time and at the latest start-up time, respectively.

A system or method, characterized in that at least some of the workflows W1.1, W2.1, W3.1 have a monitoring step of monitoring the growth of cells, in which monitoring step the degree of fusion of the cell culture is determined, and in dependence on the degree of fusion the preferred start-up time t of at least one of the following workflows W1.1, W2.2, W3.3 is determined _p And/or determining the earliest start-up time t of the following workflow W1.2, W2.2, W3.2 in the monitoring step _l And/or the latest start-up time t _l 。

A system or method characterized by a workflow W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3 comprise at least one or more of the following working steps: cell culture planting 18, counting of living or dead cells 17, cell extraction from cell culture, separation of cells from the bottom of the carrier 1 tank, addition of separating agent 14, fusion scan 11, addition of nutrients 19 or discard of cell culture 15 or seeding of cells.

A system or method, characterized in that the transport device 5 at a first time takes the first carrier 1 for executing the first workflow W1.1 from a storage location in the holding module 2 and transports it to the processing module 3 in such a way that the processing device 4 starts processing the cell culture carried by the first carrier at a first start time t, and the transport device 5 subsequently takes the second carrier 1 for executing the second workflow W2.1 from a storage location in the holding module 2 at a second time and transports it to the processing module 3 in such a way that after the execution of the first workflow W1.1 has ended, the processing device 4 starts processing the cell culture carried by the second carrier 1 at a second start time t, wherein the two start times t are calculated such that the sum Σ of the effective functions W (t) is minimized or maximized.

A system or method, characterized by a graphical data input device through which a user can pre-set the trend of an efficiency function.

A system or method, characterized in that the handling device 4 has pipetting means, microscopes and/or opening means for opening containers containing nutrients and/or that the preservation module 2 is an incubator and/or that the transport device 5 has gripping arms or end effectors and/or is automated.

A system or method, characterized in that the minimum or maximum is a local minimum or maximum and/or the minimum or maximum is in a limited number of workflows W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; values calculated over the duration of W3.1, W3.2, W3.3.

All the disclosed features are of importance for the application, as such, but also in combination with each other. The disclosure of the present application thus also includes the full disclosure of the relevant/attached priority documents (copies of the previous application) also for the purpose of incorporating the features of these documents into the claims of the present application. The dependent claims, even if they do not have the features of the claims cited, feature independent inventive developments of the prior art, in particular for the purpose of divisional applications on the basis of these claims. The application specified in each claim may additionally have one or more of the features specified in the description above, in particular provided with reference numerals and/or specified in a list of reference numerals. The application also relates to designs in which the individual features mentioned in the above description are not realized, in particular if these features are optional for the respective purpose of use or are replaced by other means having the same technical effect.

List of reference numerals

1. Carrier W1.1 workflow

2. Preservation module W1.2 workflow

3. Processing module W1.3 workflow

4. Processing device W2.1 workflow

5. Transport device W2.2 workflow

6. Control device W2.3 workflow

7. Memory device W3.1 workflow

11. Fusion scan W3.2 workflow

12. Measured fusion degree W3.3 workflow

13. Calculated preferred start-up time W1 workflow

W2 workflow

14. Work flow of adding separating agent W3

W4 workflow

15. Cell suspension W5 workflow

16. Effective function of centrifugal processing W (t)

17. Counting

18. Time to start seeding t of cells

19. Adding nutrient t _e Earliest start-up time

21. Executing workflow t _p Preferred start-up time

t _l Latest start-up time

22. Recalculating t _e 、t _p 、t _l

23. Moving sigma sum over time

A application

A1 Application of

A2 Application of

A3 Application of

Claims (14)

1. A system for culturing a biological cell culture, the system having: a plurality of carriers (1) carrying one or more cell cultures, respectively; at least one storage module (2) in which the carrier (1) is stored; a treatment module (3), in which treatment module (3) the cell culture carried by the carrier (1) is treated by means of at least one treatment device (4); a transport device (5) by means of which the carrier (1) is transported between the holding module (2) and the processing module (3); and a control device (6) having a memory device (7) and controlling the transport device (5) and the processing device (4), wherein applications (A1, A2, A3) associated with a plurality of mutually different cell cultures are stored in the memory device (7), wherein each application (A1, A2, A3) is a processing rule for processing the associated cell culture, the processing rule comprising one or more workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) which are carried out in the processing module (3) at time intervals in each case in succession at a start-up time (t), wherein one workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) contains one work step or a plurality of work steps which need to be carried out successively in a short period, wherein the control means (6) controls the simultaneous execution of a plurality of applications (A1, A2, A3) and in this case in the same processing module (3) in succession in time-series starts up the one or more workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W2.3, W3.1, W3, W3.2.3) which are functionally related to each other, W1, A2, W3.3) and the configuration of which is dependent on the one of the one workflow (A1, A2, A3, W3.3) in each case in time-series, and by minimizing or maximizing the sum (Σ) of the effective functions (W (t)), the order and start-up time (t) of the workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) of the different applications (A1, A2, A3) or of different instances of the same application are calculated.

2. Method for culturing biological cell cultures, wherein a plurality of carriers (1) each carry one or more cell cultures, the carriers (1) being stored in at least one storage module (2), wherein the cell cultures are processed in a processing module (3) by means of at least one processing device (4), wherein the carriers (1) are transported between the storage module (2) and the processing module (3) by means of a transport device (5), wherein the transport device (5) and the processing device (4) are controlled by a control device (6) having a memory device (7), and wherein applications (A1, A2, A3) assigned to a plurality of cell cultures which differ from one another are stored in the memory device (7), wherein each application (A1, A2, A3) is a processing rule for processing the assigned cell cultures, the processing rule comprising one or more workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) which are each carried out in the processing module (3) at time intervals in succession at a start-up time (t), wherein one workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) contains one working step or a plurality of working steps which need to be carried out successively in succession in a short period of time, wherein the control means (6) control a plurality of applications (A1, A2, A3 In the same processing module (3), starting the one or more workflows (W1.1, W1.2, W1.3) of the applications (A1, A2, A3) that differ from one another in a time-sequential manner; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3), characterized in that each workflow (W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3) is configured with an effective function (W (t)) related to the start time (t), and by minimizing or maximizing the sum (Σ) of the effective functions (W (t)), workflows (W1.1, W1.2, W1.3) of different applications (A1, A2, A3) or of different instances of the same application are calculated; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3) and start-up time (t).

3. The system according to claim 1 or the method according to claim 2, characterized in that each workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) is configured with a preferred start-up time (t _p ) In the case of the preferred start-up times, the workflow (W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; the effective functions (W (t)) of W3.1, W3.2, W3.3) have a minimum or maximum value.

4. A system or method according to claim 3, characterized in that at least some of the first workflows (W1.1, W1.2, W1.3) of the different applications (A1, A2, A3) have a first preferred start-up time (t _p ) The first preferred start time is within a processing duration of a second workflow, the second preferred start time (t _p ) At a first preferred start-up time (t _p ) Previously, wherein the control device (6) shifts the first and/or second activation times in time in such a way that the two first and second workflows (W1.1, W1.2, W1.3; w2.1, W2.2, W2.3; w3.1, W3.2, W3.3) are performed sequentially, wherein the first and second start-up times are selected such that the sum (Σ) of the effective functions (W (t)) is minimized or maximized.

5. A system or method according to claim 3 or 4, characterized in that at least some of the workflows (W2.1, W2.2, W2.3) are configured with an earliest start-up time (t _e ) The workflow (W2.1, W2.2, W2.3) is not allowed to be started before the earliest start-up time, and/or is configured with a latest start-up time (t _l ) After the latest start-up time, the workflow (W2.1, W2.2, W2.3) is not allowed to start, wherein it is provided in particular that the effective function (W (t)) is at the earliest start-up time (t) _e ) And latest start-upTime (t) _l ) Where has a maximum value.

6. A system or method according to any of claims 3-5, characterized in that the effective function (W (t)) has a value from the earliest start-up time (t) _e ) To the preferred start-up time (t) _p ) Descending or ascending branches and slave preferring start-up times (t _p ) To the latest start time (t _l ) Linear or non-linear rising or rising branches.

7. The system or method according to any of the preceding claims, characterized in that the trend of the effective function (W (t)) is selected such that the costs that are spent in the workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) for executing the application (A1, A2, A3) are increased in the preferred start-up time (t _p ) There is a minimum in the case of start-up and a maximum in the case of start-up of the workflow at the earliest start-up time and at the latest start-up time, respectively.

8. System or method according to any of the preceding claims, characterized in that at least some of the workflows (W1.1, W2.1, W3.1) have a monitoring step of monitoring the cell growth, in which the degree of fusion of the cell culture is determined, and in accordance with the degree of fusion the preferred start-up time (t) of at least one of the following workflows (W1.1, W2.2, W3.3) is determined _p ) And/or determining in the monitoring step an earliest start-up time (t) of the following workflow (W1.2, W2.2, W3.2) _l ) And/or the latest start-up time (t _l )。

9. The system or method according to any of the preceding claims, characterized in that the workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) comprises at least one or more of the following working steps:

storage of cell cultures (18), counting of living or dead cells (17), extraction of cells from the cell cultures, separation of cells from the bottom of the tank of the carrier (1), addition of separating agents (14), fusion scan (11), addition of nutrients (19), discarding of the cell cultures (15) or seeding of cells.

10. System or method according to any of the preceding claims, characterized in that the transport means (5) at a first time take the first carrier (1) for executing the first workflow (W1.1) from a storage location in the holding module (2) and transport it to the processing module (3) such that the processing means (4) starts processing the cell culture carried by the first carrier at a first start time (tw1.1), and that the transport means (5) then at a second time take the second carrier (1) for executing the second workflow (W2.1) from a storage location in the holding module (2) such that after the end of the execution of the first workflow (W1.1) the processing means (4) starts processing the cell culture carried by the second carrier (1) at a second start time (t), wherein both start times (t) are calculated such that the sum of the effective functions (Σ) is minimized or maximized.

11. A system or method according to any preceding claim, wherein a graphical data input device through which a user can pre-set the trend of an effective function.

12. System or method according to any of the preceding claims, characterized in that the handling device (4) has pipetting means, microscopes and/or opening means for opening containers containing nutrients and/or the holding module (2) is an incubator and/or the transporting device (5) has gripping arms or end effectors and/or is automated.

13. The system or method according to any of the preceding claims, characterized in that the minimum or maximum value is a local minimum or maximum value and/or the minimum or maximum value is a value calculated over the duration of a limited number of workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3).

14. A system or method characterised by one or more of the distinguishing features of any preceding claim.