SYSTEM FOR CULTIVATING BIOLOGICAL CELL CULTURES

Description

field of technology

[0001] The invention relates to a system or a method for cultivating biological cell cultures, comprising a plurality of carriers, each carrying one or more cell cultures, at least one storage module in which the carriers are stored, a process module in which the cell cultures carried by the carriers are treated by means of at least one treatment device, a transport device with which the carriers are transported between the storage module and the process module, and a programmed or programmable control device which has a storage device and which controls the transport device and the treatment device, wherein applications assigned to several different cell cultures are stored in the storage device, each application being a treatment instruction for treating the assigned cell culture, which comprises one or more treatment units,hereinafter referred to as workflows, which are carried out at intervals one after the other at a start time in the process module, wherein a workflow includes one or more work steps, wherein the one or more work steps must be carried out directly one after the other, wherein the control device controls the simultaneous execution of a plurality of applications and starts the one or more workflows of the different applications one after the other in a chronological order in the same process module. State of the art

[0002] Cells, particularly human cells, are grown in plastic dishes, for example in wells of carriers, by the addition of nutrients. Adherent cells are typically grown in a monolayer that adheres to the bottom of a plastic dish or a well in a carrier. Such cells are detached from the bottom before the entire bottom is covered with cells. For this purpose, a confluence scan is performed, which determines the degree of coverage of a substrate, for example the bottom of a carrier, with cells. The cells detached from the bottom can be diluted and transferred to new carriers or new wells. State-of-the-art systems and methods for maintaining such cell cultures can include an incubator, a robot for handling the carriers carrying the cells, and handling devices with which the nutrient medium can be added or changed.

[0003] US 2005 / 260743 describes a modular cell culture system and process with different treatment stations, so-called manipulation modules, using a workflow management system, also referred to as an 'automated, smart (intelligent) system'. 'Operational rules' and 'process rules' are written and stored in the software on the computer and in the programs.

[0004] EP 1 598 415 A1 describes a system for cultivating biological cell cultures, in which cells are stored on carriers in a storage module. Using a transport device comprising a gripper, the carriers can be removed from the storage module and transported to a processing module. There, the cell cultures stored on the carrier are treated by a treatment device. Different cell cultures are treated according to different treatment instructions. For each cell culture, for example, for each carrier, there can be an application that is a treatment instruction for treating the associated cell culture. Such an application can contain one or more workflows. Each workflow can contain at least one work step. However, a workflow can also comprise several work steps.The work steps of a workflow are characterized by the fact that they must be carried out immediately or in quick succession. The multiple workflows of an application, on the other hand, are each carried out at longer time intervals between individual workflows. Between the execution of individual workflows, the carriers can be located in their assigned storage locations in the storage module. The applications, which can number 100 or more, and the workflows assigned to them can be stored as digital data records in a memory device of a control device. The programmable control device coordinates the parallel execution of a plurality of applications and controls the one or more workflows of the different applications in a chronological sequence one after the other in the same process module.

[0005] An automated system for carrying out these applications, which preferably include at least one or more workflows in which the culture medium is changed or confluence is measured, is controlled by a scheduling program of the control device. Such a scheduling program creates a schedule to execute the workflows of the individual applications one after the other. The scheduling program should also be able to influence the workflows. Generally, such a schedule contains several workflows in which the duration of the workflows and the intervals between the workflows can vary unpredictably. Since cells are living entities and their properties, especially growth rates, can vary unpredictably, the workflows must be adapted to the respective individual development of the cell culture.In general, the growth rate of cells is variable and difficult to predict. For example, the growth rate of cells can vary from batch to batch due to known factors such as transfection, serum deprivation, or cryptic factors. The behavior of a cell culture, especially the growth rate, must therefore be monitored in order to adjust the timing of workflow start times to the development stage of the cell culture.

[0006] A system for cultivating biological cell cultures must also be able to carry out the applications in a time-effective manner and to define the start times of the workflows in such a way that workflows which would fall into the treatment duration of another workflow at an optimal start time are shifted in time in such a way that they are carried out one after the other in the process module. Summary of the invention

[0007] The invention is based on the object of advantageously developing a system or method for cultivating biological cell cultures in which applications are carried out simultaneously with workflows. One object, in particular, is to operate the system cost-effectively.

[0008] The problem is solved by the invention specified in the claims, wherein the subclaims are not only advantageous developments of the invention specified in claim 1, but also represent independent solutions to the problem.

[0009] A first aspect of the invention relates to avoiding temporal collisions between workflows which, under optimal conditions, would have overlapping processing times. Such workflows must be shifted in time such that the start time of one workflow occurs after the completion of the other workflow. According to the invention, each workflow is assigned an action function which is dependent on the start time. The action function or the curve of the action function can have an extremum at the time at which the workflow would preferentially be executed. The action function is thus minimal or maximum, or the curve of the action function has a minimum or maximum at a time at which the workflow would preferentially be executed. The minimum of the action function is preferred. The action function then increases at earlier times and at later times.The increase can be linear or non-linear and can increase less towards an earliest start time than towards a latest start time, or vice versa. For example, if the workflow is to monitor the condition of a cell culture, the workflow can include a step in which the degree of cell coverage of the bottom of a well in which the cells are stored is measured. Such a confluence scan can be performed using a microscope and automatic image analysis. Depending on the degree of coverage, cells can be removed in a subsequent step to reduce the coverage. Liquids can be added to detach the cells from the bottom. The cells can be removed using a pipetting device.At the same time, by measuring the coverage, a growth rate individually assigned to the cell culture can be determined by comparing the coverage of a previous measurement with the current coverage, taking into account the time elapsed in the meantime. Using this growth rate measured by the confluence scan, a future time can be calculated when the next cell culture monitoring should be carried out in a subsequent workflow. The system calculates a preferred start time. The preferred start time can, for example, be a time at which a confluence of 80% is expected based on the determined growth rate. The system can calculate a latest start time that is approximately where a confluence of 90% to 100% is predicted based on the determined growth rate. A later start time would cause the surface to overgrow.The system can calculate an earliest start time, which, for example, lies where the calculated confluence lies between 15% and 30%, preferably 20%. Using these boundary values ​​of the earliest start time, the latest start time, and the preferred start time, an action function is constructed. This action function can be normalized, for example, with the value 0 at the preferred start time and the value 1 at the earliest or latest start time. This action function can be a function of the preferred start time. However, it is also possible for such an action function to rise asymptotically slowly or rapidly to its maximum value, particularly at the latest start time, as the preferred start time approaches the earliest or latest permissible value.If the system contains applications that each have at least one workflow that, due to its preferred start time, would fall within the processing time of another workflow, the start times of the two workflows are varied in a variation calculus such that the sum of the two effect functions is minimized or has a minimum. If the system contains applications in which several workflows overlap in terms of their execution time, these workflows are subjected to a variation calculus in which the start times of the workflows are varied in such a way that the sum of all effect functions has a minimum. The minimum can be a local minimum. But it can also be a global minimum. In particular, it is intended that at least some of the workflows are assigned an earliest start time before which the workflow may not be started.However, it is also intended that at least some of the workflows have a latest start time after which the workflow may not be started. Furthermore, it can be intended that the earliest start time and the latest start time are immediately adjacent to the preferred start time, meaning that the workflow may only be started at the preferred start time. A corresponding effect function can be represented mathematically by having one or two very steep branches whose gradient increases rapidly as the function approaches the earliest or latest start value. In the simplest case, the effect functions can have linear branches. However, the branches of the effect function can also be represented by a polynomial, even of a higher degree.According to a preferred embodiment of the invention, the actual start times of the action functions for the workflows are chosen such that the sum of the action functions for the workflows is minimized, since the action function is increased if a workflow is executed at a start time other than the preferred start time. In this way, the action function can be viewed as a cost function, with the execution of a workflow too early or too late incurring a penalty or cost. The goal of the system is to minimize the sum of the action functions or cost functions across the workflows of the applications, i.e., to maximize the time efficiency of the system.

[0010] The invention also relates, among other things, to planning software or a planner for managing multiple functions with one or more workflows. In an alternative embodiment of such a planner, the functions and the goal of the planner can also be reversed. A function could be maximized at the time a workflow is started. The goal of the system would be to maximize the sum of the effective functions across the workflows of the applications in order to maximize the system's time efficiency.

[0011] A workflow can comprise at least one or more of the following steps: establishing a cell culture, counting the living or dead cells, detaching the cells from the bottom of a well on a carrier, removing cells from the cell culture to reduce the degree of coverage or confluence, confluence scanning, changing a cell culture medium, adding nutrients, or abandoning the cell culture. In particular, it can be provided that the workflows in an application are widely distributed over time, in the sense that the workflows are separated by large time intervals. For example, a workflow can last 30 minutes. During this time, a confluence scan can be performed, the cell culture medium can be changed, or nutrients can be added.The time between two consecutive workflows can be a few hours or even several days, whereby the interval between consecutive workflows depends on a growth rate of the cell culture that is constantly calculated or recalculated each time a workflow is executed. For example, in a workflow in which cell growth is monitored, a confluence scan can first be performed to determine the growth rate based on the confluence set in the previous workflow. The confluence can then be reduced to, for example, 10% by removing cells. The confluence can then be reduced to, for example, 10% by removing, separating, diluting, or seeding cells in a new carrier. Based on the determined growth rate, the preferred start time (tp ) of the following workflow can be determined such that the expected confluence is 80%.Based on the determined growth rate, the effect function is also recalculated, for example by setting an earliest start time at which the confluence is 20% and a latest start time (tl ) at which the expected confluence is 95%.

[0012] The work steps within a workflow form temporally related processes that must be performed sequentially, for example, with time intervals of ideally 0 (zero) or at most a few minutes, whereby no other workflow can be inserted between the work steps. A first work step can, for example, be the removal of a carrier from the storage module and its transport to the process module. A subsequent second work step can, for example, be a confluence scan, in which the degree of coverage of the cells adhering to the bottom of a dish is determined using optical means. A third work step can be the addition of a "detachment agent," which is used to detach the cells from the bottom of the dish or the carrier's trough.A fourth step can be the at least partial removal of the cells from the nutrient solution in the dish or well, or the transfer of a portion of the cells to a fresh dish or well. This step can be used to adjust the degree of coverage, i.e., the confluence. A fifth step can be the addition of nutrient solution. A sixth step can be the transport of the carrier back to the storage module. Each of these steps can influence the course of the active function. Further steps can be the counting of living and dead cells. This step can be performed before or after the second step. For the cultivation of a specific cell line, e.g.For CHO cells (Chinese Hamster Ovary), an instance of an application is created that contains the parameters for culturing this cell line, for example, a complete medium recipe, the confluence at which the cells are to be passed, a split ratio, and the number of dishes. An application can affect one or more dishes. An application that affects multiple dishes, each of which in turn has a multitude of dishes, wells, or carriers in which cell cultures are stored, has workflows whose length is longer than for applications that only affect one dish. The interval between two workflows depends essentially on the cell line and mainly on the doubling time or split ratio of the cells in the cell line. The split ratio specifies the degree of dilution of the cells in each workflow.Typically, the doubling time varies from cell line to cell line and thus also from instance to instance of a cell line maintenance application. There are time-critical workflows that must be performed to prevent cells from becoming overconfluent.

[0013] An application can contain workflows that vary in number and order. For example, the maintenance of iPSCs (induced pluripotent stem cells) typically includes a workflow for media changes ("feeding cells" or "refeeding cells"), since iPSCs require factors such as FGF2 that have a short half-life. Stem cells typically need to be fed every 24 hours. The frequency of a "medium change" can vary and depends on the growth rate or division ratio of the cells. The timing of confluence scans can also depend on the growth rate of the cells, so a confluence scan can, for example, be performed before or after a first or second media change.

[0014] The above statements characterize the efficiency function as a function that should be minimized if, for example, the efficiency function represents costs. However, the reverse case is also possible, for example, if the efficiency function represents a benefit. In this case, the efficiency function should be maximized. Short description of the drawings

[0015] Embodiments of the invention are explained below with reference to the accompanying drawings. They show: Fig. 1: In the upper section labeled a, three applications A1, A2, A3, each with associated workflows W1.1, W1.2, W1.3; W2.1, W2.2, W2.3 and W3.1, W3.2 and W3.3, which are to be executed at preferred start times, whereby the workflows W2.1, W2.2 and W2.3 of application A2 must be executed at fixed (unchangeable) start times. In the lower section labeled b, the workflows of the three applications are shown after they have been partially shifted in time (see arrow) such that they can be executed one after the other in a process module. Fig. 2: In the upper section labeled a, an action function W(t). The section below labeled b represents a time scale in days. The lower section labeled c represents the confluence values ​​corresponding to the time scale. Fig.3 a calculation scheme for calculating the earliest start time (te ), the preferred start time (tp ) and the latest start time (tl ), Fig. 4 a calculation scheme also for calculating the preferred start time from the result of a confluence scan, Fig. 5 a representation according to Figure 2a of an action function W(t), Fig. 6 another representation according to Figure 2a of an action function W(t), Fig. 7 an application A that contains three workflows W1, W2, W3 that are repeated cyclically, Fig. 8 an application A that contains five workflows W1, W2, W3, W4, W5 that are repeated cyclically, Fig. 9 schematically the steps for resolving a time conflict when two or more workflows are to be executed simultaneously or the start times of one workflow fall within the execution time of another workflow; Fig. 10 schematically the elements of a system according to the invention, Fig.11 schematically shows a graphical user interface for the manual design of an active function, Fig. 12 a representation according to . Figure 1 , in which two workflows W2.1 and W3.2 have been swapped in their chronological order to optimize the effect function, for example, maximize or minimize, Fig. 13 a representation according to Figure 2 , but in the case that the effect function indicates a benefit and is to be maximized. Description of the embodiments

[0016] The Figure 10 shows a schematic diagram of a system consisting of a storage module 2 for storing cell cultures. Such storage modules 2 can be incubators. The housing of such a storage module can contain several "hotels" in which a plurality of carriers 1 are stored one above the other. Regarding such storage modules 2, reference is made to WO 2020 / 098960 A1.

[0017] A transport device 5, which may comprise a gripper, end effector, and / or other transport means, for example, transport rails or an automation mechanism, is provided, with which a carrier 1 can be transported from the storage module 2 to a process module 3. Regarding the design of a process module, reference is made to WO 2020 / 098957 A1. One or more treatment devices 4 are located in the process module 3. The treatment device 4 may comprise a microscope, a pipetting device, or the like.

[0018] The preferably multiple carriers 1 stored in the storage module 2 can be microplates. Such microplates have a plurality of wells, each of which stores a nutrient medium containing cell cultures. Each carrier 1 can contain cell cultures of one cell type. Different carriers 1 can carry different cell lines. The storage module 2 is designed so that the cells stored therein proliferate at a cell-specific division rate.

[0019] A control device 6 can comprise a microcontroller or a microcomputer capable of controlling the transport device 5 and the treatment device 4. The control device 6 can further comprise a storage device 7 in which applications are stored in digital form. An application is a treatment protocol for treating cell cultures, wherein it can be provided that each carrier 1 is assigned an application, i.e. a treatment protocol, which takes into account the characteristics of the cell line carried by the carrier 1. However, an application A can also comprise multiple carriers 1, wherein it is then provided in particular that the multiple carriers 1 carry cells of the same type. Carriers 1 can be stored in the storage module 2, each of which carries different cell culture lines and to which a different application A1, A2, A3 is each assigned.Each application can have one or more workflows W1, W2, W3, W4, W5, whereby each workflow can contain one or more work steps. The workflows of an application A1, A2, A3 are to be carried out one after the other at specified time intervals, whereby each workflow W1, W2, W3 has a preferred start time at which it can be started. However, some or all of the workflows W1, W2, W3 can also be started at an earlier or later time, but not before an earliest start time te and not after a latest start time tl. The time between the workflows of an application can range from minutes to days. The preferred time between workflows is between a few hours and six days, half a day to six days, or one to four days. The duration of a workflow can be between one minute and 60 minutes.

[0020] A workflow comprises one or more work steps that must be carried out immediately or in quick succession, with a break between two consecutive work steps preferably being zero or often lasting several minutes, with no other workflow in between. A work step can, for example, be a confluence scan 11, in which a measured confluence 12 is determined in order to determine a preferred start time tp. Further work steps can be the addition of a separation agent 14, the suspension of cells 15, centrifugation, the counting of living or dead cells 17, cell priming, i.e. the seeding of new cells 18, and the addition of nutrients, i.e. the feeding of cells 19. A workflow can comprise at least one of these or another work step not previously listed.

[0021] The applications comprising one or more workflows W1, W2, W3, W4, and W5 can be executed periodically over an indefinite period of time, for example, for cell line maintenance. An application can extend over several days.

[0022] The system can have multiple instances of the same application, for example, the cell line maintenance application can have a first instance "Maintain CHO cells" or a second instance "Maintain Vero cells".

[0023] The Figure 7shows an application A consisting of three workflows W1, W2, W3, in which a first workflow W1 is carried out one to three days after the start of the application, in which a confluence scan 11 is performed. After a further one to three days, a second workflow W2 is carried out, in which a confluence scan 11 is also carried out first and, depending on the degree of confluence, the cells are washed and a detachment agent 14 is added in order to detach the cells from the bottom of a well in the carrier. While the confluence scan 11 takes about 1 minute, the subsequent step, for example washing or the addition of detachment agent 14, can take 10 to 20 minutes. In the first workflow W1, an initial confluence can be measured. With the further measurement of the confluence carried out in the second workflow W2, the division rate of the cell line can be determined.From this, the preferred start time tp of another workflow can be calculated, for example if the expected confluence is 80%.

[0024] A third workflow W3 can also be performed after five to ten minutes. This workflow can include the steps of cell collection 15, centrifugation 16, counting of viable and dead cells 17, and / or seeding cells 18, or seeding cells 18 in new microplates 18.

[0025] The Figure 8 shows another embodiment of an application in which five workflows W1, W2, W3, W4, W5 are executed one after the other and the sequence of five workflows is repeated for an indefinite period of time.

[0026] Workflow W1, workflow W2, and workflow W3 can each include a nutrient addition 19. Workflow W6 includes the confluence scan step 11. The interval between nutrient additions can be set to 24 hours, for example, and the cell doubling time can vary between cell lines, so the number of nutrient additions depends on the cell growth rate, i.e., the number of workflows in the application can vary. Depending on the start time of the confluence scan 11 and the cell growth rate, the order of workflows W1, W2, W3, and W6 can also vary (i.e., W6 can be outside the Figure 8Workflow W4 includes the confluence scan 11, and step 14 includes the removal of the medium and the addition of the detaching agent. Workflow 5 may include the suspension of the cells in step 15, centrifugation in step 16, counting of dead and live cells in step 17, and seeding of the cells in step 18.

[0027] The Figure 2 and the Figures 5 and 6 show example curves of an impact function W(t). The impact function represents a value that depends on the start time t of a workflow W1, W2, W3, W4, W5. The impact function W(t) has a minimum at a preferred start time tp of the workflow to which the impact function is assigned. The impact function has a maximum at a time corresponding to the latest start time tl or the earliest start time te.

[0028] Section b of the Figure 2gives an example of the number of days to which the start times given in section a refer. Section b of the Figure 2 Indicates the corresponding confluence, i.e., the degree of coverage of the bottom of a well in which the cell culture is stored, expressed as a percentage. The optimal start time is the time at which confluence is expected to be 80%. The latest start time is the time at which confluence is just below 100%. The earliest start time TE is the time at which confluence is expected to be approximately 20 to 30%.

[0029] The control device 6 is programmed such that, in the absence of conflict, each workflow W1, W2, W3, W4, W5 begins at the preferred start time tp. In the event of a conflict, in which two or more workflows would have to take place simultaneously according to the schedules specified by the applications A1, A2, A3, the workflows are shifted such that they can be executed one after the other in a sequence in the same process module 3. For this purpose, the workflows are shifted such that the sum of the effective functions is at a minimum. Such a shift may involve a change in the sequence, i.e., a rearrangement of the workflows.

[0030] The Figure 12 shows an example of an action function to be maximized in a representation according to Figure 2 .

[0031] The Figure 1shows an example in which three applications A1, A2, A3 each have three workflows W1.1, W1.2, W1.3; W2.1, W2.2, W2.3 and W3.1, W3.2 and W3.3. The workflows W1.1, W2.1 and W3.1 of the applications A1, A2, A3 are arranged in such a way that they would overlap if executed as scheduled. The workflows W2.1, W2.2, W2.3 of the application A2 also have the special feature that they may only be executed at the preferred start time, so the latest start time and the earliest start time are essentially identical. These workflows W2.1, W2.2 and W2.3 essentially govern the temporal ordering of the remaining workflows. A system according to the invention can thus comprise workflows W2.1, W2.2, W2.3 that govern the temporal ordering of the remaining workflows.

[0032] As a result, in order to minimize the sum of the effective functions of workflows W1.1, W2.1 and W3.1, workflow W1.1 is moved forward in time and workflow W3.1 is moved back in time.

[0033] The second workflows W2.2 and W3.2 do not need to be postponed in the example, as they can be executed sequentially as planned. Only workflow W1.2 needs to be moved forward slightly. None of the workflows W1.3, W2.3, or W3.3 need to be postponed. While the order of the workflows changes in the example shown, it is to be expected that minimizing or maximizing the sum of the effective functions may lead to a change in the order of the workflows in some cases.

[0034] The Figure 11 shows that swapping the workflows W3.2 and W2.1 is also possible.

[0035] The Figure 3shows a curve of the reproduction rate of a cell culture as a function of time. The time of the preferred start of a workflow, tp, is chosen so that confluence is expected to be 80%. The time of the latest possible start of the workflow, tl, is chosen so that confluence is expected to be 90%. The time of the earliest start of the workflow, te, is chosen so that confluence is expected to be 20%.

[0036] The Figure 4 shows the process of calculating a calculated preferred start time 13 using a confluence 12 measured at a time point during a confluence scan 11. At the time point of the confluence scan 11, a confluence of 60% was measured. The point on the growth curve at which the confluence is 80% is determined.

[0037] In this example, the system counts the resuspended cells during a previous workflow run and calculates a dilution to achieve a confluency of 10% of the cells. Based on this set 10%, the system then calculates the time at which the cells would likely be in the middle or end of a growth cycle (50 to 80% confluency). From the initial confluency of, for example, 10%, the time at which confluency is, for example, 80% is calculated, assuming an exponential growth curve.

[0038] If the system does not know the doubling time of the cells, it can conservatively use the shortest known doubling time for mammalian cells in a culture or a doubling time shorter than the shortest known doubling time. For example, the system can use a preliminary doubling time of 12 hours to calculate when to perform the confluence scan. Alternatively, the system can only perform a confluence scan every 24 hours until the cells have reached a stable doubling time. Once the system has doubling time data, it can use the cells' doubling time to determine when 80% confluence is reached.

[0039] The Figures 5 and 6 show variants of the action function where the action function W(t) has a non-linear course. Figure 5In the action function shown, the time interval between the latest start time tl and the preferred start time tp is significantly smaller than the time interval between the earliest start time te and the preferred start time tp .

[0040] The Figure 6 shows, however, a symmetrical course of the action function W(t) with respect to tp.

[0041] The Figure 11 shows as an example a graphical interface with which the course of the action function W(t) can be manually preset by "dragging" anchor points on a screen.

[0042] It may be desirable for users to add applications, workflows, or other work processes to the system, which can present another challenging scheduling problem for the system to handle. Times within a schedule can be changed. In particular, it can be taken into account that some cell lines grow faster or slower than expected. During a workflow, one or more carrier plates can be treated with the treatment device 4 in immediate succession. This is preferably done using the same work steps.

[0043] According to the invention, a cell culture system is provided. In one implementation, the cell culture system has an incubator, a refrigerator, two storage areas for laboratory equipment at room temperature, and a process module 3. The process module 3 can have a liquid handler, for example a pipette. Other mechanisms, such as a decapper for decapping bottles, a gripper for handling carriers 1, as well as a microscope, can be provided. Depending on the applications and workflows or other work processes, the capacity of the system can be limited either by the capacity of the incubator for the carriers, in particular carrier plates, and in particular the number of plates the incubator can process, or by the capacity (speed) of the process module. Thus, only a limited number of workflows is possible in a process module 3.

[0044] The system can run applications that, for example, involve cell line maintenance or a cell-based assay or experiment. These applications can run for an indefinite period of time with respect to cell line maintenance, particularly extending over days or weeks.

[0045] The workflows include, in particular, the confluence scan, the addition of a detachment agent, and cell collection. The cell collection workflow can contain individual operations, namely cell collection and cell seeding. Furthermore, work steps such as centrifugation or the counting of dead and live cells can be included. For the cultivation of a specific cell line, e.g., CHO cells, an instance of the application is created that contains the parameters for culturing this cell line. Parameters can include the complete medium recipe, passage confluence, the split ratio, or the number of carriers, for example, carrier plates such as microplates.

[0046] The work steps within a workflow are connected and have no gaps, and in particular no gaps of more than one minute between two work steps. The time between two workflows in the same application is greater than one minute and is typically minutes, hours, or days. The transport of a carrier from the storage module 2 to the process module 3 and its treatment there can be delayed if the required confluence has not yet been achieved. A workflow can therefore contain work steps that are performed optionally, for example, if a confluence scan reaches a certain value. If the certain value is not reached in the confluence scan, one or more subsequent work steps can be omitted or postponed.

[0047] The method underlying the system according to the invention is capable of processing a schedule in which the duration and intervals of the workflows differ from each other. The method can be described as follows with reference to the Figure 9 summarize:

[0048] Defining a workflow to a preferred start point tp , an earliest start time te and a latest start time tl as well as the costs for executing the workflow at a start time other than tp , where the costs are represented in the action function W(t) (Execution of Workflow 21).

[0049] If a user input is made or if data is found that would, for example, change the preferred start time of a subsequent workflow, the data (start times) of all subsequent workflows are recalculated (22). The calculation is performed in such a way that a sum, in particular a total sum of the effective functions of the workflows, reaches a minimum (23). The minimum does not have to be a global minimum; it can be a local minimum.

[0050] The sum Σ of all effect functions W(t) of all workflows can be represented as a multidimensional function. Σ = W 1 t 1 + W 2 t 2 + … + W n t n

[0051] From which a global or local minimum can be calculated by shifting the respective start times and taking into account the duration of each workflow d dt Σ = 0 which is subject to the constraint that all workflows follow one another in time and the start time of each of the workflows does not lie within the time period of any other workflow.

[0052] The invention provides a method for mixing the workflows in which a sum of action functions W(t) is minimized, which represents, for example, the costs to be incurred for the implementation of the workflows, so that the method according to the invention is a cost-optimized method.

[0053] The invention thus relates to a planner or planning software with which a plurality of applications can be managed, each application containing one or more workflows. The workflows of the various applications are executed in an optimized manner, using the optimization characteristics described above.

[0054] The above statements serve to explain the inventions covered by the application as a whole, which each independently develop the state of the art by at least the following combinations of features, whereby two, several or all of these combinations of features can also be combined, namely:

[0055] A system characterized in that each workflow W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3 is assigned an action function W(t) dependent on the start time t and the sequence and the start times (t) of the workflows W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3 of different applications A1, A2, A3 are calculated by minimizing the sum Σ of the action functions W(t).

[0056] 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 assigned an effect function W(t) dependent on the start time t and the order and the start times (t) of the workflows W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3 of different applications A1, A2, A3 or instances of the same application are calculated by minimizing the sum Σ of the effect functions W(t).

[0057] A system or 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 assigned a preferred start time tp at which the action function W(t) assigned to the workflow W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3 has a minimum.

[0058] A system or method characterized in that at least some first workflows W1.1, W1.2, W1.3 of the various applications A1, A2, A3 have a first preferred start time tp which falls within a treatment duration of a second workflow whose second preferred start time tp lies before the first preferred start time tp, wherein the control device 6 shifts the first and / or the second start 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 carried out one after the other, wherein the first and second start times are selected such that the sum Σ of the action functions W(t) is minimized or maximized.

[0059] A system or method characterized in that at least some of the workflows W2.1, W2.2, W2.3 are assigned an earliest start time te before which the workflow W.1, W2.2, W2.3 may not be permitted and / or a latest start time tl is assigned after which the workflow W2.1, W2.2, W2.3 may not be permitted, wherein it is provided in particular that the effect function W(t) has a maximum at the earliest start time te and at the latest start time tl.

[0060] A system or method characterized in that the action function W(t) has a branch which decreases or increases linearly or non-linearly from an earliest start time te to the preferred start time tp and a branch which increases or decreases linearly or non-linearly from the preferred start time tp to the latest start time tl.

[0061] A system or method characterized in that the course of the action function W(t) is selected such that the costs to be expended for the execution of the application A1, A2, A3 at the start of the workflows W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3 have a minimum at the preferred start times tp and a maximum at the earliest and latest start times.

[0062] A system or method characterized in that at least some of the workflows W1.1, W2.1, W3.1 have a cell growth monitoring step in which the confluence of a cell culture is determined, and depending thereon the preferred start time tp of at least one subsequent workflow W1.1, W2.2, W3.3 is determined and / or that in the monitoring step the earliest start time tl and / or the latest start time tl of a subsequent workflow W1.2, W2.2, W3.2 is determined.

[0063] A system or method 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 work steps: establishing a cell culture 18, counting the living or dead cells 17, removing cells from the cell culture, detaching the cells from the bottom of a well of the carrier 1, adding a separation agent 14, confluence scan 11, adding nutrients 19 or abandoning the cell culture 15 or seeding cells.

[0064] A system or method characterized in that the transport device 5 removes a first carrier 1 for carrying out a first workflow W1.1 at a first time from a storage location in the storage module 2 and transports it to the process module 3 such that the treatment device 4 begins to treat the cell culture carried by the first carrier at a first start time t and the transport device 5 subsequently removes a second carrier 1 for carrying out a second workflow 2.1 at a second time from a storage location in the storage module 2 and transports it to the process module 3 such that the treatment device 4 begins to treat the cell culture carried by the second carrier 1 after completion of the execution of the first workflow W1.1 at a second start time t, wherein the two start times t are calculated such that the sum Σ of the action functions W(t) is minimized or maximized.

[0065] A system or method characterized by a graphical data input device that allows a user to specify the course of an action function.

[0066] A system or a method which is characterized in that the treatment device 4 has a pipetting device, a microscope and / or an opening device for opening containers containing nutrients and / or that the storage module 2 is an incubator and / or that the transport device 5 has a gripper arm or end effector and / or is automated.

[0067] A system or method characterized in that the minimum or maximum is a local minimum or maximum and / or that the minimum or maximum 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.

[0068] All disclosed features are essential to the invention (individually, but also in combination with one another). The disclosure of the application hereby fully incorporates the disclosure content of the associated / attached priority documents (copy of the prior application), also for the purpose of incorporating features of these documents into claims of the present application. The subclaims, even without the features of a referenced claim, characterize independent inventive developments of the prior art with their features, in particular for filing divisional applications based on these claims. The invention specified in each claim may additionally comprise one or more of the features provided in the above description, in particular with reference numerals, and / or specified in the list of reference numerals.The invention also relates to designs in which individual features mentioned in the above description are not implemented, in particular insofar as they are clearly unnecessary for the respective intended use or can be replaced by other technically equivalent means. List of reference symbols 1 carrier W1.1 Workflow 2 Storage module W1.2 Workflow 3 Process module W1.3 Workflow 4 Treatment facility W2.1 Workflow 5 Transport device W2.2 Workflow 6 Control device W2.3 Workflow 7 Storage device W3.1 Workflow 11 Confluence scan W3.2 Workflow 12 measured confluence W3.3 Workflow 13 calculated preferred start time W1 Workflow W2 Workflow 14 Adding a release agent W3 Workflow W4 Workflow 15 Suspending the cells W5 Workflow 16 Centrifuge W(t) Active function 17 To count 18 Cell seeding t Start time 19 Nutrient addition te earliest start time 21 Executing a workflow tp preferred start time t l latest start time 22 Recalculate te , tp , tl 23 temporal postponement ∑ sum A application A1 application A2 application A3 application

Claims

1. A method for cultivating biological cell cultures, in which a plurality of carriers (1) each carry one or several cell cultures, which are stored in at least one storage module (2), wherein the cell cultures are treated in a process module (3) by means of at least one treatment device (4), wherein the carriers (1) are transported with a transport device (5) between the storage module (2) and the process module (3), wherein a control device (6) having a memory device (7) is used to control the transport device (5) and the treatment device (4), and applications (A1, A2, A3) assigned to different cell cultures are stored in the memory device (7), wherein each application (A1, A2, A3) is a treatment specification for treating the assigned cell culture, which contains one or several workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3), which are performed at time intervals successively at a respective start time (t) in the process module (3), wherein a workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) contains one work step or several work steps to be performed one after the other at short notice, wherein the control device (6) controls the simultaneous performance of a plurality of applications (A1, A2, A3), and thereby starts one or several 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) in chronological sequence one after the other in the same process module (3), characterized in that each workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) has assigned to it an active function W(t) that depends on the start time (t), and the sequence and start times (t) of the workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) of different applications (A1, A2, A3) or instances of the same application are calculated by minimizing or maximizing the sum (Σ) of active functions (W(t)).

2. The method according to claim 1, characterized in that each workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) has assigned to it a preferred start time (tp), at which the active function (W(t)) assigned to the workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) has a minimum or maximum.

3. The method according to claim 2, characterized in that at least some first workflows (W1.1, W1.2, W1.3) of the different applications (A1, A2, A3) have a first preferred start time (tp), which fall within a treatment duration of a second workflow whose second preferred start time (tp) lies before the first preferred start time (tp), wherein the control device (6) shifts the first and / or second start 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 one after the other, wherein the first and second start times are selected in such a way as to minimize or maximize the sum (Σ) of active functions (W(t)).

4. The method according to one of claims 2 or 3, characterized in that at least some of the workflows (W2.1, W2.2, W2.3) have allocated to them an earliest start time (te) before which the workflow (W2.1, W2.2, W2.3) must not be started and / or a latest start time (tl) after which the workflow (W2.1, W2.2, W2.3) must not be started, wherein it is provided in particular that the active function (W(t)) has a maximum at the earliest start time (te) and at the latest start time (tl).

5. The method according to one of claims 2 to 4, characterized in that the active function (W(t)) has a branch that linearly or nonlinearly descends or ascends from an earliest start time (te) to the preferred start time (tp) and a branch that linearly or nonlinearly ascends or descends from the preferred start time (tp) to the latest start time (tl).

6. The method according to one of the preceding claims, characterized in that the progression of the active function (W(t)) is selected in such a way that the costs to be expended for performing the application (A1, A2, A3) at the start of the workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) have a minimum at the preferred start times (tp) and a respective maximum at the earliest and latest start times.

7. The method according to one of the preceding claims, characterized in that at least some of the workflows (W1.1, W2.1, W3.1) have a monitoring step that monitors cell growth, in which the confluence of a cell culture is determined, and the preferred start time (tp) of at least one ensuing workflow (W1.1, W2.2, W3.3) is determined depending thereupon, and / or that the earliest start time (te) and / or the latest start time (tl)of an ensuing workflow (W1.2, W2.2, W3.2) is determined in the monitoring step.

8. The method according to one 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 several of the following work steps: Creating a cell culture (18), counting the living or dead cells (17), removing cells of the cell culture, detaching the cells from the bottom of a trough of the carrier (1), adding a detaching agent (14), confluence scan (11), adding nutrient (19), discontinuing the cell culture (15) or seeding cells.

9. A system for cultivating biological cell cultures, with a plurality of carriers (1), which each carry one or several cell cultures, with at least one storage module (2), in which the carriers (1) are stored, with a process module (3), in which the cell cultures carried by the carriers (1) are treated by means of at least one treatment device (4), with a transport device (5), with which the carriers (1) are transported between the storage module (2) and the process module (3), and with a control device (6), which has a memory device (7) and which controls the transport device (5) and the treatment device (4), wherein applications (A1, A2, A3) assigned to several different cell cultures are stored in the memory device (7), wherein each application (A1, A2, A3) is a treatment specification for treating the assigned cell culture, which contains one or several workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3), which are performed at time intervals successively at a respective start time (t) in the process module (3), wherein a workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) contains one work step or several work steps to be performed one after the other at short notice, wherein the control device (6) controls the simultaneous performance of a plurality of applications (A1, A2, A3), and thereby starts one or several 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) in chronological sequence one after the other in the same process module (3), characterized in that each workflow (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) has assigned to it an active function W(t) that depends on the start time (t), and the sequence and start times (t) of the workflows (W1.1, W1.2, W1.3; W2.1, W2.2, W2.3; W3.1, W3.2, W3.3) of different applications (A1, A2, A3) or instances of the same application are calculated by minimizing or maximizing the sum (Σ) of active functions (W(t)).

10. The system according to claim 9, characterized in that the control device 6 is programmed to implement a method according to one of claims 1 to 8.

11. The system according to claim 9 or 10, characterized in that the transport device (5) removes a first carrier (1) for performing a first workflow (W1.1) from a storage space in the storage module (2) at a first time and transports it to the process module (3) in such a way that the treatment device (4) begins to treat the cell culture carried by the first carrier at a first start time (tW1.1), and the transport device (5) then removes a second carrier (1) for performing a second workflow (W2.1) from a storage space in the storage module (2) at a second time and transports it to the process module (3) in such a way that the treatment device (4) begins to treat the cell culture carried by the second carrier (1) at a second start time (t) after the performance of the first workflow (W1.1) has concluded, wherein the two start times (t) are calculated in such a way as to minimize or maximize the sum (Σ) of the active functions (W(t)).

12. The system according to one of claims 9 to 11, characterized by a graphic data input device, with which a user can set the progression of an active function.

13. The system according to one of claims 9 to 12, characterized in that the treatment device (4) has a pipetting device, a microscope and / or an opening device for opening nutrient-containing containers, and / or that the storage module (2) is an incubator, and / or that the transport device (5) has a gripping arm or end effector and / or is automated.

14. The system according to one of claims 9 to 13, characterized in that the minimum or maximum is a local minimum or maximum, and / or that the minimum or maximum 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).

15. Software for a system according to one of claims 9 to 14, which is programmed to implement a method according to one of claims 1 to 8.