Continuous method and improved device for the analysis of cancer samples, kit and use

Abstract

The present invention in one aspect thereof relates to a laboratory automation device that is adapted to support an automated and continuous processing method for identifying an anti-cancer drug or anti-cancer drug combination for patients that suffer from, or are being diagnosed for, a neoplastic disease or tumor. Provided are furthermore a respective automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination as well as kits and respective uses thereof.

Description

[0001] CONTINUOUS METHOD AND IMPROVED DEVICE FOR THE ANALYSIS OF CANCER SAMPLES, KIT AND USE

[0002] The present invention in one aspect thereof relates to a laboratory automation device that is adapted to support an automated and continuous processing method for identifying an anti-cancer drug or anti-cancer drug combination for patients that suffer from, or are being diagnosed for, a neoplastic disease or tumor. Provided are furthermore a respective automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination as well as kits and respective uses thereof.

[0003] BACKGROUND OF THE INVENTION

[0004] Generally, drugs in development for therapeutic purposes are screened for efficacy in a conventional screening system, where drugs from a library are tested in a suitable cellbased assay. Usually, the viability of the cells and / or the cytotoxicity of the candidate drug are investigated. This approach often is done in a so-called high throughput, but still there is a high risk that drugs identified as promising in such approach will later disappoint in the subsequent clinical testing.

[0005] Furthermore, it has turned out that in specific fields of indication, drug combinations are increasingly desired, e.g., to avoid the development of resistances, or to exploit synergistic effects.

[0006] Also, the predictability of early-stage experiments to the future in vivo situation needs to be improved, in order to reduce the risk of failure of drug combinations that have turned out promising in the preclinic.

[0007] So far, almost no systematic approaches have been disclosed to screen potential drug combinations at early stage. Conventionally, drugs are combined empirically, by medical practitioners, and tested in patients. However, a systematic approach to really investigate the combinatorial effects of such drug combination is missing. This means that a huge potential of promising drug combination exists but never sees the patient, for lack of systematic investigation.

[0008] In common lab-automation solutions, these are set up in order to support a batch-wise assay processing, comprising the steps of experiment planning and respective preparations, followed by the start of the system as used, and loading of the batch of samples. This is followed by the assay conduct and the respective analysis. Then, the batch of samples is removed, and an experiment cleanup is performed, requiring a stop or pause of the system stop, before a new experiment can be started.

[0009] WO 2022 / 008044A1 relates to a laboratory automation device. Furthermore, the invention relates to a method, a computer program, a computer-readable medium and a controller for operating a laboratory automation device. The device comprises a door of a housing for accessing a workspace, wherein the door comprises a transparent display for displaying information and for allowing a person to view into the workspace.

[0010] An inherent characteristic cell-based screening is that current systems can only start screening en bloc. In most cases, the individual models are not ready at the same time. This becomes even more difficult when the screens are being run with patients or other sources that are not ready at the same time but need to be run without interruption. Current automated screening approaches work by starting a large batch, running it to completion and then starting the next batch when it is finished. However, to accommodate clinically relevant response analysis, longer assay times are required to cover a wide range of different mechanisms of action and to provide outcome analysis that can be translated directly into the clinic (in vitro clinical trial).

[0011] The testing of complex in vitro models requires longer assay times and therefore a higher flexibility when to start new drug testing processes within a fully automated testing device is needed. Furthermore, in an experimental setting using fresh samples of patients, tissue samples are ready at variable points in time. This is also the case in research settings, where samples may be equally ready for processing at different timepoints, e.g. from in vitro cell culture experiments. A flexible system that allows to incorporate a new test sample (loading) into the analysis system while running an otherwise continuous analysis process would ensure the fastest handling in that the samples can be loaded whenever ready. At the same time, prevention of any contamination is of utmost importance for continuous operation and inter office handling differences need to be excluded as well. A fast handling of samples also ensures a better biological representation of the data as generated and provides a higher data quality. Furthermore, no storage of samples and tissues is required, and any addition of preservatives which could affect sample quality and results.

[0012] Therefore, it is an object of the invention to provide new and improved methods and devices in order to analyze the phenotype and / or genotype of cells obtained from tissue samples upon exposure of these cells to drugs or drug combinations, in particular in order to streamline and render more effective patient-specific treatment approaches.

[0013] The present invention generally relates to the automated screening of drugs or combinations thereof, in particular patient-specific drugs or combinations thereof, for therapeutic purposes. In particular, the present invention relates to the automated screening of drugs or combinations thereof that can be used therapeutically in the treatment and / or prevention and the slowing down of the progression of neoplastic or tumorous cell growth in an individual in need thereof.

[0014] In a first aspect, the present invention solves the above object by providing a laboratory automation device for an automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination, the device comprising i) at least one workspace area enclosed by at least one housing; ii) at least one automated handling arm for handling samples, drug samples, and / or cell culture materials, iii) optionally, at least one tissue sample dissociation unit for dissociating patient-derived cancer cell comprising tissue samples in order to obtain dissociated cancer cells, iv) at least one unit for producing at least one array of 3D microtissues, such as microtumors, based on cancer cells, such as the dissociated cancer cells of step ii), v) at least one drug testing unit for contacting the at least one array of said 3D microtissues with at least one potential anti-cancer drugs and / or combinations thereof, vi) at least one first analysis unit for determining an effect of said drug and / or combinations of drugs on the at least one array of said 3D microtissues as produced in step iv), vii) at least one second analysis unit for identifying an anti-cancer drug or anti-cancer drug or combination based on the effect as determined in step vi), and optionally further comprising a unit for selecting said anti-cancer drug or anti-cancer drug combination as identified, and viii) the housing comprising at least one loading door comprising a loading system suitable for a sterile loading and unloading of patient samples. The loading door is preferably designed as an airlock system with interconnected door that prevent direct connection between the inside and outside of the system.

[0015] Preferred is the device according to the present invention, further comprising a second loading door comprising a loading and unloading system for a sterile feeding or removing of cell culture consumable materials.

[0016] In the context of the present invention, the term “sterile” shall mean a minimal biological contamination of an environment, such as a surface, medium or device, including, but not limited to, perfect sterility.

[0017] Further preferred is the device according to the present invention, wherein the workspace area comprises 1, 2, 3, 4 or 5 of the units in one housing, preferably, wherein preferably all units except the at least one second analysis unit are enclosed in one housing.

[0018] Preferred is the device according to the present invention, wherein any separately housed units are interconnected with suitable doors and / or locks providing an uninterrupted and sterile loading and unloading of patient samples and / or materials between the units.

[0019] Further preferred is the device according to the present invention, further comprising means to automatically detect and remove contaminations, in particular comprising means for machine learning-based recognition of contaminations, such as fungi, and / or means for image trend analysis-based recognition of contamination, such as bacteria.

[0020] In a second aspect, the present invention solves the above object by providing a kit compatible with the loading door comprising a loading system of the device according to the present invention, wherein the kit comprises all reagents as required for performing a complete assay cycle for one patient sample. Preferred is such reagent kit to provide an outer containment to the reagents and materials as mechanical protection and sterile barrier. It contains measures or structures to hold the reagents and materials in place inside the kit and it may also protect its contents from measures for decontamination (e.g. UV light) during sterile loading / unloading.

[0021] Preferred is a kit containment that contains design structures that prevent incorrect loading of the kit and assure secure closure of the containment outside of the testing device (e.g. locking mechanism). Furthermore, the kit is designed to allow a continuous operation of the device, e.g. using suitable compartmentalization.

[0022] Preferred is the kit according to the present invention, comprising an environmental monitoring tag for tracking the transport and handling conditions, wherein the monitoring tag optionally comprises means for wireless communication, such as, for example, an RFID.

[0023] Further preferred is the kit according to the present invention, comprising a unique identifier, such as, for example a barcode or RFID, that is manually or automatically read upon loading of the kit in order to ensure a correct assignment of the sample and to identify the content thereof.

[0024] In a third aspect, the present invention solves the above object by providing a continuous and automated method for identifying anticancer drugs or drug combinations for patients that suffer from, or are being diagnosed for, a neoplastic disease or tumor, the method comprising a) providing a laboratory automation device according to the present invention, b) providing at least one first sample, such as patient-derived sample comprising cancer cells, to the laboratory automation device according to step a), and starting a first identification cycle at starting timepoint Tl, wherein the first identification cycle comprises the steps of i) optionally, dissociating the cancer cells of the sample of step b) a in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on said (dissociated) cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drug and / or combination of drugs, iv) determining an anti-cancer effect of the drug and / or combinations thereof of step iii) on said array of said 3D microtissues, and iv) identifying an anticancer drug or drug combination based on the effect as determined in step iii) at timepoint T2, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified; c) providing at least one second sample, such as a patient-derived sample comprising cancer cells, to the laboratory automation device according to step a) through the loading system thereof, and starting a second identification cycle at starting timepoint T3, wherein timepoint T3 is after T1 and before T4, wherein the second identification cycle comprises the steps of i) optionally, dissociating the cancer cells of the sample of step b), in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on said (dissociated) cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drug and / or a combination of drugs, iv) determining an anticancer effect of the drug and / or combinations thereof of step iii) on said array of said 3D microtissues, and iv) identifying an anticancer drug or drug combination based on the effect as determined in step iii) at timepoint T4, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified.

[0025] Preferred is the method according to the present invention, wherein the first sample comprising cancer cells is provided to the laboratory automation device according to step a) through the loading system thereof.

[0026] Further preferred is the method according to the present invention, wherein the anticancer drug or drug combination is furthermore identified as patient-specific and / or personalized.

[0027] Preferred is the method according to the present invention, furthermore comprising the step of storing the results of the identifying step on a suitable storage medium, such as a computer.

[0028] Further preferred is the method according to the present invention, wherein 2, 3, 4, 5, 6, or more independent identification cycles are performed, preferably at different starting timepoints Tx, wherein X is an integer corresponding to the beginning of the independent identification cycle as performed. In a fourth aspect, the present invention solves the above object by providing the use of the device according to the present invention or the kit according to the present invention in an automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination according to the method according to the present invention.

[0029] The present invention provides devices and methods to continuously analyze tissue biopsy samples of a patient or preselected group of patients that suffer from, or are diagnosed for, a neoplastic disease or tumor. The samples are used to generate 3D microtissues, in particular so-called “microtumors” that are then used in testing for effective and optimally patient-specific effective therapeutic drugs of combinations of drugs for a treatment and / or the prevention of said neoplastic disease or tumor. The methods and systems allow for a continuous and thus faster analysis and search for drugs, and in particular screens for drugs and drug combinations that are effective for the actual patient and / or specific patient-group. A flexible system that allows to incorporate a new test sample (loading) into the analysis system while running an otherwise continuous analysis process ensure furthermore the fastest handling in that the samples can be loaded whenever ready. A fast handling of samples also ensures a better biological representation of the data as generated and provides a higher data quality. Furthermore, no storage of samples and tissues is required, and any addition of preservatives which could affect sample quality and results.

[0030] As mentioned above, the present invention provides a laboratory automation device for an automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination as described herein. The device comprises several components in order to allow for a testing according to the present invention, such as i) at least one workspace area enclosed by at least one housing; ii) at least one automated handling arm for handling samples, drug samples, and / or cell culture materials, iii) optionally, at least one tissue sample dissociation unit for dissociating patient-derived cancer cell comprising tissue samples in order to obtain dissociated cancer cells, iv) at least one unit for producing at least one array of 3D microtissues, such as microtumors, based on cancer cell samples, such as the dissociated cancer cells of step ii), v) at least one drug testing unit for contacting the at least one array of said 3D microtissues with at least one potential anti- cancer drug and / or combinations thereof, vi) at least one first analysis unit for determining an effect of said drug and / or combinations thereof on the at least one array of said 3D microtissues as produced in step iv), vii) at least one second analysis unit for identifying an anti-cancer drug or anti-cancer drug combination based on the effect as determined in step vi), and optionally further comprising a unit for selecting said anti-cancer drug or anti-cancer drug combination as identified. Importantly, the device furthermore provides the feature that viii) the housing comprises at least one loading door comprising a loading system suitable for a sterile loading and unloading of samples, such as patient samples.

[0031] The at least one loading door comprising a loading system suitable for a sterile loading and unloading of patient samples can be made from common materials, like plastic, metal, aluminum, steel or glass. Preferably, the materials are chosen in order to allow decontamination as well as heating and / or cooling of the door and the loading system and its components.

[0032] In the context of the present invention, the inventive device is therefore designed in order to allow an efficient and continuous processing of samples to be analyzed. Furthermore, the device is substantially, preferably fully, automated in order to require only minimal interaction of the user with the processes.

[0033] The advantages of the system are achieved by providing a combination of features that allow for a continuous running of the device. This is first and foremost achieved by providing one or more loading door(s) each comprising a loading (and / or unloading) system suitable for a sterile loading and unloading of patient samples as well as consumables and / or process waste, such as spent media and even contaminations. When providing additional openings in the device or system (in this case, the loading door(s)), at the same time the system has to be designed to allow a continuing processing of samples as already processed in the device, and furthermore an efficient, non-interruptive and sterile integration of additional samples to be analyzed in parallel. This issue also occurs in between separate housings inside the overall device, if provided accordingly (see below for different layouts). Preferred is therefore a device according to the present invention, wherein any separately housed units are interconnected with suitable doors and / or locks providing an uninterrupted and sterile loading and unloading of patient samples and / or materials between the units.

[0034] EP4019130A1 discloses a door for a laboratory workstation, said laboratory workstation comprising a horizontally extending work surface for providing a working area thereon, in particular a bench. The door is configured to be movable in relation to the work surface in a vertical direction perpendicular to the work surface between a safety position, in which the door closes the working area for manual access of a user, an access position in which the working area is open for manual access of a user, and a loading position. The concept of the loading door is different from the system of the present invention, moving the door from the safety position into the loading position allows limited, manual access of a user to the working area or parts thereof through an access opening. Furthermore, a loading or unloading of the work surface through the access opening may occur, while the one or more process operation is paused. That is, a manual access is still possible, either fully (access position) or limited (loading position). No loading system is provided, and the operation of the machine has to stop while loading.

[0035] The device according to the present invention can be integrated, i.e. the workspace area may comprise 1, 2, 3, 4 or 5 of the units in one housing, wherein preferably all units except the at least one second analysis unit are enclosed by one housing.

[0036] The loading system may be provided with a lock system for a sterile loading of materials or consumables as used in the system(s) and / or unloading waste and / or products as produced in the system(s). The lock system can be attached to or is integrated into the system, for example as an integral part in a housing.

[0037] Further preferred is the device according to the present invention that further comprises a second loading door comprising a loading and unloading system for a sterile feeding or removing of cell culture consumable materials, consumables and / or process waste as described herein. This allows for separate and simultaneous loading and / or unloading of materials and products in or from the system.

[0038] Further preferred is the system according to the present invention, wherein said lock system has doors, flaps and / or hatches to open and close the system arranged on each end of the lock, in particular on each opposing end thereof. In general, the lock system works in an in / out fashion, the outside flap or door or hatch is opened, materials are loaded into the lock space (optimally in form of a tray or box, e.g. as described below), and the outside flap or door or hatch is closed. Then, the inside flap or door or hatch is opened, either by a respective mechanism, or by the robot arm as forming part of the inside system, and the materials are forwarded into the device space, and stored or used. When unloading materials from the system, the inside flap or door or hatch is opened either by a respective mechanism, or by the robot arm, materials, such as products or consumables / waste are loaded into the lock space (optimally in form of a tray or box, e.g. as described below), and the inside flap or door or hatch is closed. Then, the outside flap or door or hatch is opened, and the materials can be taken out of the system. Preferred is the system according to the present invention, wherein said doors, flaps and / or hatches are for opening or closing reciprocally. This ensures that the system is not open to the outside at any given time of the loading and / or unloading step.

[0039] Preferred is the device according to the present invention, wherein the loading door comprising the loading system comprises an inner and outer door, wherein the outer door can be locked, in particular automatically, in order to prevent user manipulation and contamination.

[0040] Further preferred is the device according to the present invention, wherein said loading system is adapted to specifically fit to a transport box or container, preferably as described herein, wherein further preferably said transport box or container comprises at least one door to be opened and closed inside the device. This allows a closed transport of products or materials in or out of the device or between devices or parts / units thereof. The transport box or container can be used for storage as well, in particular when insulated. The transport box or container can be made from common materials, like plastic, metal, aluminum, steel or glass. The box can have handles for handling, and / or holders for labeling.

[0041] When using a kit, box or container adapted to fit with the present device (see below), the device according to the present invention may include a loading system (or systems) comprising means for unlocking the cover and opening of the kit, box or container upon insertion thereof.

[0042] Advantageously, in a preferred device according to the present invention, the loading system further comprises means for sterilizing the materials as loaded and / or unloaded, such as, for example, a lock system for sterilizing using UV, and / or wherein said lock system further comprises means for thawing or cooling / freezing the materials to be loaded or unloaded. In this embodiment, the materials (and / or) the box or tray(s) are sterilized before entering or leaving the system. During the sterilization, flaps or doors or hatches of the lock system are preferably both closed, in order to avoid UV irradiation or gases (ozone) to leave the lock, and to ensure an effective sterilization to an essential extent. The doors and the means for sterilizing can include a control unit, optimally including a timer, either as integral part of the device or as a separate device.

[0043] In the device according to the present invention, the loading system can further comprise means selected from UV decontamination means or a HEPA filtered air exchange for decontaminating the system after a loading or unloading, decontaminating a container cover, such as a kit cover, after loading, and / or decontaminating a contaminated test plate, if necessary.

[0044] Further preferred is the device according to the present invention, wherein said lock system further comprises means for thawing or cooling / freezing the materials to be loaded or unloaded. In this embodiment, the materials (and / or) the box or tray(s) are heated or cooled before entering or leaving the system. During the heating or cooling, the flaps or doors or hatches are preferably both closed, in order to maintain the desired temperature(s), and to ensure an effective heating or cooling to an essential extent. The doors, flaps or hatches and the means for heating or cooling can include a control unit, optimally including a timer, either as integral part of the system or as a separate device. The lock can also include separate temperature zones, like 4°C and -20° for cooling / thawing and freezing or 37°C and ambient. The temperature zones can include a control unit which allows a gradual control of the heating and cooling, e.g. from 4°C to - 20°C or from 4°C to 37°C (all temperatures are about the value as given).

[0045] Further preferred is the device according to the present invention, wherein the loading system comprises a tracking system, such as, for example an RFID or barcode reader.

[0046] Even further preferred is the device according to the present invention, wherein the optional tissue sample dissociation unit and said unit for producing an array of 3D microtissues are positioned in the same housing, and / or wherein said drug testing unit and said first analysis unit are positioned in the same housing, and preferably wherein said two housings are interconnected to form a discrete device and / or wherein said device is, at least in part, arranged vertically.

[0047] Another important aspect of the device according to the present invention relates to the autonomous organization and functioning thereof. Mechanically, the device according to the present invention may comprise at least one automated handling arm for handling cell and tissue samples, drug samples, and / or cell culture materials that is a robotic arm comprising suitable means for gripping samples, drug samples, and / or cell culture materials.

[0048] Furthermore, the device according to the present invention preferably comprises implemented computer program means for automatically and autonomously operating, monitoring, organizing and prioritizing all assay steps, in order to ensure an uninterrupted and automated execution of all device activities.

[0049] Preferably, in the device according to the present invention, the means for organizing further provide a self-organized prioritization of critical process steps and timing constraints, such as, for example, incubation times, and / or means for predicting and prenotification of a user to prevent waste overspills, consumables shortages and / or device interruptions.

[0050] Particularly preferred is a “dynamic prioritization” as provided by the program means, wherein certain process steps or events are prioritized over others in order to ensure defined process conditions of process steps with the highest priorization(s). Also, the dynamic prioritization of samples provides a flexible order of assays and the steps thereof for a maximum capacity and efficiency of the device. In case of colliding activities and process steps, the computer program means may also autonomously postpone or even omit uncritical or low priority process steps in order to assure correct execution of critical or high priority process steps.

[0051] Furthermore, the controlling computer program means may include a self-recovery means, e.g. after a power failure or interruption. This allows to recover all running processes and continue these, if interrupted. For example, the imaging step may be repeated.

[0052] Also preferred is the device according to the present invention, further comprising computer program means to ensure a correct assignment of a kit, box or container as used to a patient sample, and / or a logging of all device activities.

[0053] As mentioned above, in common lab-automation solutions, these are set up in order to support a batch-wise assay processing, comprising the steps of experiment planning and respective preparations, followed by the start of the system as used, and loading of the batch of samples. This is followed by the assay conduct and the respective analysis. Then, the batch of samples is removed, and an experiment cleanup is performed, requiring a stop of the system stop, before a new experiment can be started.

[0054] In contrast, the device according to the present invention runs continuously and automated with no system stop at any time when samples are or will be analyzed, except for device maintenance. Advantageously, the device autonomously organizes all processing tasks in order to ensure uninterrupted assay execution. Furthermore, the inventive device is designed to allow continuous operation with no user interaction required within the inner workspace / housing. The inventive device is furthermore designed to prevent, minimize and reduce contamination and has active measures to detect and remove contaminated tests (or reagents).

[0055] The device according to the invention meets the requirements of complex in vitro model testings that require longer assay times and therefore a higher flexibility when to start new drug testing processes / cycles within an otherwise fully automated testing device. Continuous loading ensures that the patient samples can be loaded and analyzed whenever ready.

[0056] Another aspect of the device according to the present invention relates to the avoidance and removal of contamination(s). Preferred is therefore a device according to the present invention, further comprising means to automatically detect, monitor and remove contaminations, in particular comprising means for machine learning-based recognition of contaminations, such as fungi, and / or means for image trend analysis-based recognition of contamination, or algorithms, such as 2D Fourier transformation for a detection of e.g. bacteria. Further preferred is the device according to the present invention, wherein the means provide an automatic decontamination of the device during idle times.

[0057] For detecting and removing contaminated plates or samples from the device, the invention provides a machine learning approach for discrete object detection, such as filamentous fungi, large microorganisms that individually visible, e.g. yeasts, and / or chemical precipitations, such as crystals. In addition, or alternatively, an image parameter trending approach is provided in order to detect small microorganisms, e.g. bacteria, that affect the turbidity of cell culture media (see, for example, Strutt JPB, et al. Machine learning-based detection of adventitious microbes in T-cell therapy cultures using long-read sequencing. Microbiol Spectr. 2023 Aug 30; 1 l(5):e0135023. doi: 10.1128 / spectrum.01350-23. Epub ahead of print. PMID: 37646508; PMCID: PMC10580871; Zhang J, et al. A comprehensive review of image analysis methods for microorganism counting from classical image processing to deep learning approaches. Artif Intell Rev. 2022;55(4):2875-2944. doi: 10.1007 / sl0462-021-10082-4. Epub 2021 Sep 29. PMID: 34602697; PMCID: PMC8478609). Preferred is a system that discriminates static objects (e.g. foreign particles, scratches etc.) against chemical (e.g. crystals) or biological contaminations.

[0058] In the context of the present invention, a “3D microtissue” shall designate an in vitro generated cell aggregate comprising cells as desired. Consequently, a microtumor shall mean a 3D microtissue generated from, at least in part, selected cancer cells derived from a cell line or a neoplastic sample, such as a tumor (see, for example, Rimann et al., An in vitro osteosarcoma 3D microtissue model for drug development, Volume 189, 10 November 2014, Pages 129-135). These two terms herein are used interchangeably.

[0059] In the context of the present invention, an “array” is a set of separate 3D microtissues that tested / analyzed, such as, for example, 3D microtumors in a multi-well plate. An array comprises at least 2 or more, or 3, 4, 5, 6, 7, 8, 9, 10 or more, 12, 48, 96, 128, 384 or more, or even 200, 300, 400, or more 3D microtumors / microtissues.

[0060] The other technical components of the device according to the present invention can be generally taken from the art, such as, for example, from WO 2021 / 110799A2 (hereby incorporated by reference in its entirety), for example a device according to the present invention, wherein the tissue sample dissociation unit comprises at least one of i) a pipetting unit, ii) an enzyme reservoir, iii) a reservoir for cell culture media, iv) a reservoir for washing solutions, v) optionally, an ultrasonic device, and vi) a centrifuge unit, and / or wherein said unit for producing an array of 3D microtissues, such as microtumors, based on said dissociated cells comprises at least one of i) a pipetting unit, ii) a cell counting unit, and, iii) a handler for microtiter plates, and / or wherein said drug testing unit comprises at least one of i) a handler for microtiter plates, ii) a pipetting unit, iii) a reservoir for cell culture media, iv) an array of reservoirs comprising at least two different drugs or combinations thereof, and iv) an incubator unit, and / or wherein said first and / or second analysis unit comprises i) a handler for microtiter plates, and / or ii) an imaging system comprising a microscope and a camera, and optionally a high resoltion (HR) scanner. Optionally, said system can further comprise a reporting unit displaying, storing, saving information on the results.

[0061] Preferred is a device according to the present invention, further comprising a unit for selecting said patient-specific drug or drug combination as identified.

[0062] Preferred is a device according to the present invention, wherein said tissue sample dissociation unit comprises at least one of i) a pipetting unit, ii) an enzyme reservoir, iii) a reservoir for cell culture media, iv) a reservoir for washing solutions, v) optionally, an ultrasonic device, and vi) a centrifuge unit.

[0063] Preferred is a device according to the present invention, wherein said unit for producing an array of 3D microtissues, such as microtumors, based on said dissociated cells comprises at least one of i) a pipetting unit, ii) a cell counting unit, and, iii) a handler for microtiter plates.

[0064] Preferred is a device according to the present invention, wherein said drug testing unit comprises at least one of i) a handler for microtiter plates, ii) a pipetting unit, iii) a reservoir for cell culture media, iv) an array of reservoirs comprising at least two different drugs or combinations thereof, and iv) an incubator unit.

[0065] Further preferred is a device according to the present invention, wherein said first and / or second analysis unit comprises i) a handler for microtiter plates, and / or ii) an imaging system comprising a microscope and a camera, and optionally an HR scanner.

[0066] Further preferred is the device according to the present invention, wherein said tissue sample dissociation unit and said unit for the production of an array of 3D microtissues share the same pipetting unit and / or wherein said drug testing unit and said first analysis unit share the same handler for microtiter plates. Even further preferred is a device according to the present invention, wherein said tissue sample dissociation unit and said unit for the production of an array of 3D microtissues share the same pipetting unit. This provides a particularly compact device. Even further preferred is a device according to the present invention, wherein said drug testing unit and said first analysis unit share the same handler for microtiter plates. This provides a particularly compact device.

[0067] Even further preferred is a device according to the present invention, wherein said tissue sample dissociation unit and said unit for producing an array of 3D microtissues are positioned in the same housing. This provides a particularly compact device. Even further preferred is a device according to the present invention, wherein said drug testing unit and said first analysis unit are positioned in the same housing. This provides a particularly compact device. Even further preferred is a device according to the present invention, wherein the two housings are connected to form a discrete system. Preferably, this includes that the housings are connected into a single device.

[0068] In another aspect of the device according to the present invention, the device can be sterilized as a whole or in parts thereof and / or comprise means for establishing and / or maintaining sterile conditions, such as sterilized by UV irradiation, ozone treatment, radiation, and / or includes sections with laminar flow, etc.

[0069] In another important aspect of the system according to the present invention, the device can be arranged vertically, either as a whole or in parts / units thereof. This provides a particularly compact device, which is particularly suitable in a point of care setting.

[0070] In the context of the present invention, the patient samples or cell culture consumable materials can be selected from any suitable biological materials, such as, for example, selected from the group consisting of microtumors, biopsy material, digestion enzymes, culture medium, cells, cancer cells, supporting cells, control cells, cancer-type specific maintenance medium, drugs, and prefabricated drug matrices.

[0071] As used herein, the term “sample” refers to a clinical sample obtainable or obtained from a patient suspected to have a neoplastic disease, e.g., a tumor / cancer, or a patient with confirmed neoplastic disease, e.g., a tumor / cancer. The sample may be obtained by any known type of biopsy, e.g., needle biopsy, liquid biopsy, core biopsy, tumor resection, needle aspiration, surgical removal of solid neoplastic tissue, as known in the art. The sample may comprise a cell sample and / or a tissue sample.

[0072] Another important aspect of the present invention then relates to a kit, box or container that is adapted and thus compatible with the loading door comprising a loading system of the device according to the present invention.

[0073] In brief, a kit, box or container according to the present invention contains all necessary reagents to run a complete assay of a sample as described herein. These include media, microtumors, biopsy material, digestion enzymes, culture medium, cells, cancer cells, supporting cells, control cells, cancer-type specific maintenance medium, drugs, and prefabricated drug matrices.

[0074] Preferably, the kit, box or container according to the present invention comprises a UV- resistant cover to allow UV decontamination in the loading dock. The cover of the kit, box or container according to the present invention contains a stacking plate to keep contents in place and to separate contents from cover to prevent sticking in frozen state. The kit cover is preferably locked to a base plate to ensure secure transport and no contamination (also in the frozen state).

[0075] Preferred is a kit containment that contains design structures that prevent incorrect loading of the kit and assure secure closure of the containment outside of the testing device (e.g. locking mechanism). Furthermore, the kit is designed to allow a continuous operation of the device, e.g. using suitable compartmentalization.

[0076] Upon insertion into the loading dock, the cover lock is mechanically unlocked, and the cover is opened upon opening of the inner dock door (see above). The kit, box or container according to the present invention may be equipped with an environmental monitoring tag to track transport and handling conditions, and this monitoring tag may communicate wireless (e.g. an RFID). Advantageously, the kit, box or container according to the present invention may contain a unique identifier (e.g. a barcode, an RFID) that is manually or automatically read upon loading in order to ensure a correct assignment to sample and to identify its content.

[0077] Therefore, preferred is a kit, box or container that is adapted and thus compatible with the loading door comprising a loading system of the device according to the present invention, wherein the kit, box or container comprises all reagents as required for performing a complete assay cycle for one selected patient sample.

[0078] As mentioned above, when using a kit, box or container adapted to fit with the present device, the device according to the present invention may include a loading system (or systems) comprising means for unlocking the cover and opening of the kit, box or container upon insertion thereof.

[0079] Therefore, preferred is a kit, box or container according to the present invention, comprising a unique identifier, such as, for example a barcode or RFID, that is manually or automatically read upon loading of the kit in order to ensure a correct assignment of the sample and to identify the content thereof. Further preferred is a kit, box or container according to the present invention comprising a UV-resistant cover for allowing a UV decontamination inside the loading system.

[0080] Further preferred is a kit, box or container according to the present invention wherein the lock is mechanically unlocked upon insertion into the loading system and the cover is opened upon opening of an inner loading system door.

[0081] The kit, box or container according to the present invention can be made from common materials, like plastic, metal, aluminum, steel or glass. Preferably, the materials are chosen in order to allow sterilization as well as heating and / or cooling of the kit, box or container according to the present invention and its components. For decontamination within the device, the kit material protects the contained materials / reagents of the kit from the effects of decontamination.

[0082] Another important aspect of the present invention then relates to a device according to the present invention, comprising at least one kit, box or container according to the present invention, e.g. present inside or inserted the housing(s) of the device.

[0083] Another important aspect of the present invention then relates to a continuous and automated method for identifying anticancer drugs or drug combinations for patients that suffer from, or are being diagnosed for, a neoplastic disease or tumor, the method comprising a) providing a laboratory automation device according to the present invention as described herein, b) providing at least one first sample, such as a patient- derived sample, comprising cancer cells to the laboratory automation device according to step a), and starting a first identification cycle at starting timepoint Tl, wherein the first identification cycle comprises the steps of i) optionally dissociating the cancer cells of the sample of step b) a in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on said dissociated cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drugs and / or combinations thereof, iv) determining an anti-cancer effect of the drug and / or combination of drugs of step iii) on said array of said 3D microtissues, and iv) identifying an anticancer drug or drug combination based on the effect as determined in step iii) at timepoint T2, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified; c) providing at least one second sample, such as a patient-derived sample, comprising cancer cells to the laboratory automation device according to step a) through the loading system thereof, and starting a second identification cycle at starting timepoint T3, wherein timepoint T3 is after T1 and before T4, wherein the second identification cycle comprises the steps of i) optionally, dissociating the cancer cells of the sample of step b), in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on said dissociated cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drug and / or combination of drugs, iv) determining an anti -cancer effect of the drug and / or combination of drugs of step iii) on said array of said 3D microtissues, and iv) identifying an anticancer drug or drug combination based on the effect as determined in step iii) at timepoint T4, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified.

[0084] Preferred is the method according to the present invention, wherein the first sample such as patient-derived sample, comprising cancer cells is provided to the laboratory automation device according to step a) through the loading system thereof.

[0085] Further preferred is the method according to the present invention, wherein the anticancer drug or drug combination is furthermore identified as patient-specific and / or personalized. Preferred is a method according to the invention that further comprises the step of selecting said patient-specific drug or drug combination as identified, i.e. isolating a certain set of drugs as patient-specific, be it as a dataset or even by physically providing a patient-specific cocktail of drugs.

[0086] Preferred is the method according to the present invention, further comprising the step of storing the results of the identifying step on a suitable storage medium, such as a computer, which may be included in the device and / or is remote, e.g. connected through a network or the internet.

[0087] In this aspect, the present invention provides methods to continuously analyzes cells or tissue biopsy samples of a patient or preselected group of patients that suffer from, or are diagnosed for, a neoplastic disease or tumor. The samples are used to generate 3D microtissues, in particular so-called “microtumors” that are then used in testing for effective and optimally patient-specific effective therapeutic drugs of combinations of drugs for a treatment and / or the prevention of said neoplastic disease or tumor. The methods allow for a continuous and thus faster analysis and search for drugs, and in particular screens for drugs and drug combinations that are effective for the actual patient and / or specific patient-group.

[0088] Specific and advantageous features of the method according to the present invention are in particular relating to the step(s) of providing at least one second sample, such as a patient-derived sample, comprising cancer cells to the laboratory automation device according to the present invention through the at least one loading system as provided.

[0089] This allows for the overall method to enable a continuous loading and continuous processing of fresh cellular (living) samples. Other features then relate to the method comprising a software as implemented to automatically operate, monitor, organize and prioritize all testing activities, and a self-organized prioritization of critical process steps and timing constraints (e.g. incubation times) based on the process according to the invention consisting of tasks made of different steps with different priorities. The automatic device activities enable uninterrupted operation of the method according to the present invention, and hence the method itself. The method includes a management of concurrent task execution, preventing conflicts.

[0090] Furthermore, a contamination detection of the method allows automatic elimination of contaminated samples and may include machine learning-based recognition of contaminations (e.g. fungus), and / or image trend analysis-based recognition of contamination (e.g. bacteria), with subsequent automatic decontamination, isolation and export of contaminated samples, as required. Furthermore, the method may include the step of an automatic decontamination of device during idle times.

[0091] The method furthermore predicts and pre-notifies a user to prevent waste overspills, consumables shortages or device interruptions. The method comprising the software as implemented further ensures the correct assignment of kits, boxes or containers as traveling through the loading / unloading system to samples, and logging of all activities.

[0092] In continuous operation, the loading dock(s) provide(s) for process-independent sample loading, and in-situ decontamination of loaded materials / consumable kits, if required. The device may include interconnected doors and locks for uninterrupted and sterile loading / unloading.

[0093] Similar to what is described above with respect to the device, the method may comprise a software as implemented, further ensuring a continuous 24 / 365 operation of the device apart from rare device maintenance interruptions, e.g. in a yearly interval.

[0094] New samples and kits, boxes or containers can be added to the process anytime as long as capacity is available, and the loading system / dock is free.

[0095] At the same time, the method comprising the software prioritizes and automatically schedules assays tasks. For this, as described above, tasks are assigned different priority levels and timing constraints, wherein higher-priority tasks preempt lower-priority ones.

[0096] The real time responsiveness of the methods and the devices according to the present invention are designed to meet the demands of real-time systems, ensuring timely task execution, while observing a load balance in that tasks are evenly distributed across available resources in order to prevent overload and thus to optimize performance.

[0097] The method and system according to the present invention is highly adaptable and adjusts (automatically) to changing device and system conditions through re-allocating resources as needed.

[0098] The method comprising the software as implemented assures the optimal state of the device by automatically detecting and eliminating contaminations, such as contaminated test tubes, cultures or plates again as described above. It furthermore provides a timely notification of a user to replace / empty waste and system materials / consumables.

[0099] Finally, the method comprising the software as implemented handles errors automatically to minimize requirement for user intervention.

[0100] As mentioned, the method according to the present invention provides a continuous automated and thus faster analysis method, and in particularly screens faster for drugs and drug combinations that are effective for the actual patient and / or specific patient- group as analyzed.

[0101] Preferred is the method according to the present invention, wherein 2, 3, 4, 5, 6, or more independent identification cycles are performed. The cycles are overlapping, and start at different starting timepoints Tx, wherein X is an integer, such as an odd integer, corresponding to the beginning of the independent identification cycle as performed. Depending on the capacity of the device(s) as used and the availability of samples, for example, a first identification cycle starts at starting timepoint Tl. Of course, the cycle may include several samples that are analyzed in parallel, if the materials are available at the same time. The samples and materials are preferably loaded into the device in the form of a sample kit through the loading door and lock system. Once the first cycle has started, the device runs continuously and automatically as described herein. After some time, at starting timepoint T3, the second identification cycle starts by loading the second sample(s) and materials into the device in the form of a sample kit through the loading door and lock system. In addition, 2, 3, 4, 5, 6, or more additional independent identification cycles can be started at starting timepoints T5, T7, T9, Til, and T13 respectively. Again, this depends on the capacity of the device(s) as used and the availability of samples. As described herein, the data as generated is collected and handled automatically, and the functionality of the device is monitored as well. The identification cycles in this example will end at timepoints T2, T4, T6, T8, T10, T12, and T14, respectively.

[0102] Therefore, preferred is the method according to the present invention, further comprising a cycle-independent sample loading through a loading system, such as, for example a door and lock system as described herein.

[0103] As mentioned above, the method according to the present invention may further comprise the step of an automated prediction and pre-notifi cation of a user to prevent waste overspills, consumables shortages or device interruptions, and / or may further comprise a self-organized prioritization of critical process steps and timing constraints by the device, such as, for example cycle incubation times.

[0104] Preferably, the method according to the present invention may further comprise the step of detecting contamination and an automatic elimination of contaminated samples, preferably further comprising an isolation and export of the contaminated samples. The detection may comprise a machine learning-based recognition of contaminations, such as, for example, fungi and / or an image trend analysis-based recognition of contaminations, such as, for example, bacteria. The method according to the present invention may further comprise an in-situ decontamination of materials as loaded through a / the loading system.

[0105] The method according to the present invention may further comprise the step of an automatic decontamination of the at least one device during idle times before, during or after performing the method. Decontamination can be performed as described herein, e.g. using UV radiation as provided by respective units as included in the device and / or lock system.

[0106] Many general steps of the method according to the present invention can be generally taken from the art, such as, for example, from WO 2021 / 110799A2 (hereby incorporated by reference in its entirety), for example for the method according to the present invention, wherein dissociating said tissue sample comprises i) if required, dissecting said tissue sample into smaller pieces comprising cells, ii) treating said tissue sample with a solution comprising at least one enzyme capable of dissociating cells in said tissue sample, preferably at least one enzyme selected from a protease, a collagenase, trypsin, elastase, hyaluronidase, papain, chymotrypsin, deoxyribonuclease I, and neutral protease (dispase), producing a supernatant comprising dissociated cells, and ii) removing said supernatant comprises said dissociated cells and suitably collecting said cells, wherein steps (ii) and (iii) are repeated at least once, wherein preferably before step (ii) said tissue sample is sonicated with ultrasound, wherein the energy of said ultrasound is set at a suitable level to not destroy a substantial amount of said cells. Preferably, in the method according to the present invention, step b) may comprise adding or removing stroma cells, stromal fibroblasts, endothelial cells and immune cells to said dissociated cells, and / or wherein in step b) for each 3D microtissue a predetermined number of cells is provided, such as, for example, between 500 and 10000 viable cells, and / or wherein in step b) said 3D microtissues are generated in at least one system selected from a hanging drop system, and b) a multiwell system, preferably comprising Ultra Low Adherence (ULA) wells. The addition or removal of cells helps controlling the microtissue environment. Particularly preferred is the addition of immune cells that are known to interact with tumors and cells in the tumor and / or are known to invade tumors in vivo, e.g. so-called tumor-infiltrating immune cells. Examples are PBMCs, lymphocytes, recombinant T cells, and the like. These settings and strategies are of particular advantage for tests and assay in the context of cancer-immunotherapy, and can furthermore involve the addition of other factors, like cytokines etc. to the microtissue environment. Both cells and factors can be added also after the formation of the mnicrotissue(s).

[0107] Preferably, in the method according to the present invention, the generation of said 3D microtissues does not require the use of a solubilized basement membrane preparation, like Matrigel®. This has the advantage that the complete method can be performed at temperatures above 4°C, such as room temperature. The generation of said 3D microtissues may comprise self-assembly of said cells comprised in said dissociated cells, and / or wherein the generation of said 3D microtissues comprises a maturation time of about 6 hours to 7 days, preferably about 1 to 6 days, more preferably about 2 to 5 days, and / or wherein said 3D microtissues as generated have a size of 350 pm + / - 100 pm. Desired and preferred is a size that is suitable for a proper analysis, in particular for the optical analysis methods as disclosed herein.

[0108] Further preferred is the method according to the present invention, wherein said contacting in step c) comprises a continuous exposure to said at least two drugs and / or combinations thereof, and / or an exposure to and subsequent removal to said at least two drugs and / or combinations thereof, optionally for at least one, preferably two and more cycles. In the method according to the present invention, the determining of said effect in step d) may be selected from size determination of said 3D microtissue, quantification of internal reporter gene expression in said 3D microtissue, determination of the intracellular ATP content in said 3D microtissue, and determination of pre-selected biomarkers in said 3D microtissue, wherein preferably said size determination of said 3D microtissue comprises at least one parameter selected from diameter, perimeter, volume, and area of optical cross section, and wherein preferably said size determination of said 3D microtissue comprises the use of an imaging device, and optionally further comprising the analysis of growthkinetics. Growth kinetics is an autocatalytic reaction which implies that the rate of growth is directly proportional to the concentration of cell. The cell concentration can be measured by direct and indirect methods that are known to person of skill and described in the literature (e.g. cell count).

[0109] In yet another aspect of the method according to the present invention, the size determination of said 3D microtissue comprises the use of an optical method, such as using an imaging device. Particularly useful is high-resolution scanning electron microscopy (HR-SEM).

[0110] The sample, such as the patient-derived tissue sample, may be selected from any suitable sample comprising cells to be used in the present assay methods, and is preferably selected from a sub-sample derived from a primary tissue sample, a primary tumor sample, and a metastasis sample, and wherein preferably said tissue sample has been obtained by a method comprising core biopsy, tumor resection, liquid biopsy and / or needle aspiration, and / or wherein said tissue sample and / or the dissociated cells are frozen and re-thawed prior to the generation of said 3D microtissues. Preferred is a method according to the present invention, wherein said tissue sample and / or the dissociated cells are frozen and re-thawed prior to the generation of said 3D microtissues.

[0111] A variant of the method according to the present invention comprises providing a primary tissue sample, obtaining a subsample in addition to the patient-derived sample and subjecting said subsample to at least one of molecular profiling, histological analysis, and histochemical analysis. In this aspect, in parallel to the 3D microtissues additional patient- or tumor-specific information can be gathered by “splitting” the material at the start of the analysis, such as, for example, patient anamnesis, hereditary information regarding the patient, and patient genomic information.

[0112] Another aspect of the present invention then relates to the use of the device according to the present invention or the kit according to the present invention in an automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination according to the method according to the present invention as described herein.

[0113] Another aspect then relates to method of treating a patient that suffers from, or is being diagnosed for, a neoplastic disease or tumor, comprising performing the method according to the present invention, and the step of suitably treating said patient with a patient-specific, in particular personalized, drug or drug combination as identified.

[0114] Preferred is a method according to the present invention, wherein said method provides a recommendation with regard to a suitable patient- or patient-group-specific drug or drug combination in order to more effectively treat the patient suffering from, or being diagnosed for, a given neoplastic disease or tumor.

[0115] In another aspect of the method according to the present invention, the drugs or combinations to which the 3D microtissues are contacted with in step c) are selected from the group consisting of cytotoxic, cytostatic and / or chemotherapeutic agents, targeted drugs, immunotherapeutic agents and / or combinations thereof. Examples for cytotoxic, cytostatic and / or chemotherapeutic agents are Anastrozole, Azathioprine, Beg, Bicalutamide, Chloramphenicol, Ciclosporin, Cidofovir, Coal tar containing products, Colchicine, Danazol, Diethylstilbestrol, Dinoprostone, Dithranol containing products, Dutasteride, Estradiol, Exemestane, Finasteride, Flutamide, Ganciclovir, Gonadotrophin, chorionic, Goserelin, Interferon containing products (including peg-interferon), Leflunomide, Letrozole, Leuprorelin acetate, Medroxyprogesterone, Megestrol, Menotropins, Mifepristone, Mycophenolate mofetil, Nafarelin, Oestrogen containing products, Oxytocin (including syntocinon and syntometrine), Podophyllyn, Progesterone containing products, Raloxifene, Ribavarin, Sirolimus, Streptozocin, Tacrolimus, Tamoxifen, Testosterone, Thalidomide, Toremifene, Trifluridine, Triptorelin, Valganciclovir, and Zidovudine. Targeted drugs are medications that increase in concentration in some parts of the body relative to others, such as antibodies. Examples are in brain cancer: bevacizumab, everolimus; breast cancer: bevacizumab, everolimus, lapatinib, pertuzumab, trastuzumab and its antibody drug conjugates; in colorectal cancer: aflibercept, bevacizumab, cetuximab, panitumumab, regorafenib, and dermatofibrosarcoma protuberans: imatinib. Immunotherapeutic agents are used in immunotherapy that is a form of cancer treatment that uses the power of the body's immune system to prevent, control, and eliminate cancer. Examples are monoclonal antibodies to treat cancer, CAR T-cell therapy, immune checkpoint inhibitors to treat cancer, cancer vaccines, immunomodulating drugs (IMiDs), and cytokines.

[0116] In yet another aspect of the method according to the present invention, said method further comprises the step of generating a database comprising information generated based on steps a) to e) and / or comprising information with respect to said patient-specific effect and / or the efficacy of a drug or drug combination for the treatment of a given neoplastic disease or tumor. Preferably, this comprises adding additional data into said database selected from i) data about the molecular profile of said tissue sample, ii) results of a histological analysis of said tissue sample, iii) results of a histochemical analysis of said tissue sample, iv) patient anamnesis, e.g. selected from the group comprising checking / determining vital functions, patient history, smoking habits, sports activities, hormone levels, previous or current medications, etc., v) hereditary information regarding the patient, and vi) genomic information for the patient.

[0117] Another important aspect of the present invention then relates to a method for loading and / or unloading of materials into the system according to the invention using the lock system according to present invention or a kit, box or container according to present invention as disclosed above. The method for loading and / or unloading of materials into the system according to the invention may be combined with the methods as disclosed herein regarding the testing(s) of compounds and cells.

[0118] The terms “of the (present) invention”, “in accordance with the invention”, “according to the present invention” and the like, as used herein are intended to refer to all aspects and embodiments of the invention described and / or claimed herein. The terms “about” and “approximately” denote an interval of accuracy that the person skilled in the art will understand to still ensure the technical effect of the feature in question. The term typically indicates deviation from the indicated numerical value by ±10%, and for example ±5%. As will be appreciated by the person of ordinary skill, the specific such deviation for a numerical value for a given technical effect will depend on the nature of the technical effect. For example, a natural or biological technical effect may generally have a larger such deviation than one for a man-made or engineering technical effect. As will be appreciated by the person of ordinary skill, the specific such deviation for a numerical value for a given technical effect will depend on the nature of the technical effect. For example, a natural or biological technical effect may generally have a larger such deviation than one for a man-made or engineering technical effect. Where an indefinite or definite article is used when referring to a singular noun, e.g. “a”, “an” or “the”, this includes a plural of that noun unless something else is specifically stated.

[0119] All references, patents, and publications cited herein are hereby incorporated by reference in their entirety.

[0120] As used herein, the term “neoplastic” refers to any growth of tissue, which has lost growth control, including solid or liquid tumors, warts, metastatic growth, etc. As used herein, the term “tumor” refers to any benign or malignant tissue in any given organ system or tissue, for example, liver, kidney, brain, breast, prostate, skin, etc.

[0121] As used herein, the term “microplate” refers to any arrangement of multiple cavities that can be used as reaction vessels or culture vessels, for example 24 well plates, 48 well plates, 96 well plates, 384 well plates, etc. as known in the art. It is also contemplated to use and arrangement of individual vessels, which do not necessarily have to be arranged as a plate, but may also be arranged informal strips of individual vessels, etc.

[0122] As used herein, the term “sonification” relates to the treatment of a tissue or cells with ultrasound to thereby destroy the integrity of certain target structures such as red blood cells in a process called hemolysis. This permits the removal from the tissue sample of hemoglobin, which can disturb the analysis of the obtained cells, particularly the optical analysis of cells due to interference of the hemoglobin with the detection means. Hemolysis can also be achieved by incubation of tissue in solutions of very high or very low osmolality inducing the burst or shrinkage of the red blood cells, by treatment with certain hemolytic enzymes, etc.

[0123] As used herein, the term “patient” refers to a human or non-human patient. The nonhuman patient may preferably be a mammal, for example, a horse, a dog, a cat, etc., that have been diagnosed with or are suspected to have a neoplastic disorder / cancer.

[0124] As used herein, the term “drug” refers to any active agent, small molecule or biotechnologically-produced molecule or combination of molecules, nucleic acid construct s), e.g. a vector for gene therapy, the effect of which shall be tested in vitro using the herein described methods and means.

[0125] The present invention relates to the following items.

[0126] Item 1. A laboratory automation device for an automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination, the device comprising i) at least one workspace area enclosed by at least one housing; ii) at least one automated handling arm for handling samples, drug samples, and / or cell culture materials, iii) optionally, at least one tissue sample dissociation unit for dissociating patient-derived cancer cell comprising tissue samples in order to obtain dissociated cancer cells, iv) at least one unit for producing at least one array of 3D microtissues, such as microtumors, based on cancer cells, such as the dissociated cancer cells of step ii), v) at least one drug testing unit for contacting the at least one array of said 3D microtissues with at least one potential anti-cancer drug and / or combination of drugs, vi) at least one first analysis unit for determining an effect of said drug and / or combinations thereof on the at least one array of said 3D microtissues as produced in step iv), vii) at least one second analysis unit for identifying an anti-cancer drug or anti-cancer drug combination based on the effect as determined in step vi), and optionally further comprising a unit for selecting said anticancer drug or anti-cancer drug combination as identified, and viii) the housing comprising at least one loading door comprising a loading system suitable for a sterile loading and unloading of patient samples.

[0127] Item 2. The device according to Item 1, further comprising a second loading door comprising a loading and unloading system for a sterile feeding or removing of cell culture consumable materials.

[0128] Item 3. The device according to Item 1 or 2, wherein said loading system is adapted to specifically fit to a transport box or container, wherein preferably said transport box or container comprises at least one door to be opened and closed inside the system.

[0129] Item 4. The device according to any one of Items 1 to 3, wherein the workspace area comprises 1, 2, 3, 4 or 5 of the units in one housing, preferably, wherein preferably all units except the at least one second analysis unit are enclosed in one housing.

[0130] Item 5. The device according to any one of Items 1 to 4, wherein any separately housed units are interconnected with suitable doors and / or locks providing an uninterrupted and sterile loading and unloading of patient samples and / or materials between the units.

[0131] Item 6. The device according to any one of Items 1 to 5, wherein said tissue sample dissociation unit and said unit for producing an array of 3D microtissues are positioned in the same housing, and / or wherein said drug testing unit and said first analysis unit are positioned in the same housing, and preferably wherein said two housings are interconnected to form a discrete device and / or wherein said device is, at least in part, arranged vertically.

[0132] Item 7. The device according to any one of Items 1 to 6, wherein the loading system further comprises means for sterilizing the materials, such as, for example, a lock system for sterilizing using UV, and / or wherein said lock system further comprises means for thawing or cooling / freezing the materials to be loaded or unloaded.

[0133] Item 8. The device according to any one of Items 1 to 7, further comprising implemented computer program means for automatically operating, monitoring, organizing and prioritizing all assay steps for the sample(s), in order to ensure an uninterrupted and most effective execution of all device activities.

[0134] Item 9. The device according to Item 8, wherein the means for organizing further provide a self-organized prioritization of critical process steps and timing constraints, such as, for example, incubation times, and / or means for predicting and pre-notification of a user to prevent waste overspills, consumables shortages and / or device interruptions.

[0135] Item 10. The device according to any one of Items 1 to 9, further comprising means to automatically detect and remove contaminations, in particular comprising means for machine learning-based recognition of contaminations, such as fungi, and / or means for image trend analysis-based recognition of contamination, such as bacteria.

[0136] Item 11. The device according to Item 10, wherein the means provide an automatic decontamination of the device during idle times.

[0137] Item 12. The device according to any one of Items 1 to 11, further comprising computer program means to ensure a correct assignment of a kit, box or container as used to a patient sample, and / or a logging of all device activities.

[0138] Item 13. The device according to any one of Items 1 to 12, wherein the at least one automated handling arm for handling samples, drug samples, and / or cell culture materials is a robotic arm comprising suitable means for gripping samples, drug samples, and / or cell culture materials.

[0139] Item 14. The device according to any one of Items 1 to 13, wherein said tissue sample dissociation unit comprises at least one of i) a pipetting unit, ii) an enzyme reservoir, iii) a reservoir for cell culture media, iv) a reservoir for washing solutions, v) optionally, an ultrasonic device, and vi) a centrifuge unit, and / or wherein said unit for producing an array of 3D microtissues, such as microtumors, based on said dissociated cells comprises at least one of i) a pipetting unit, ii) a cell counting unit, and, iii) a handler for microtiter plates, and / or wherein said drug testing unit comprises at least one of i) a handler for microtiter plates, ii) a pipetting unit, iii) a reservoir for cell culture media, iv) an array of reservoirs comprising at least two different drugs or combinations thereof, and iv) an incubator unit, and / or wherein said first and / or second analysis unit comprises i) a handler for microtiter plates, and / or ii) an imaging system comprising a microscope and a camera, and optionally an HR scanner. Item 15. The device according to Item 14, wherein said tissue sample dissociation unit and said unit for the production of an array of 3D microtissues share the same pipetting unit and / or wherein said drug testing unit and said first analysis unit share the same handler for microtiter plates.

[0140] Item 16. The device according to any one of Items 1 to 15, wherein the loading door comprising the loading system comprises an inner and outer door, wherein the outer door can be locked, in particular automatically, in order to prevent user manipulation and contamination.

[0141] Item 17. The device according to any one of Items 1 to 16, wherein the loading system comprises means selected from UV decontamination means or a HEPA filtered air exchange for decontaminating the system after a loading or unloading, decontaminating a container cover, such as a kit cover, after loading, and / or decontaminating a contaminated test plate, if necessary.

[0142] Item 18. The device according to any one of Items 1 to 17, wherein the loading system comprises means for unlocking the cover and opening of a kit upon insertion thereof.

[0143] Item 19. The device according to any one of Items 1 to 18, wherein the loading system comprises a tracking system, such as, for example an RFID or barcode reader.

[0144] Item 20. The device according to any one of Items 1 to 19, wherein the patient samples or cell culture consumable materials are selected from the group consisting of microtumors, biopsy material, digestion enzymes, culture medium, cells, cancer cells, supporting cells, control cells, cancer-type specific maintenance medium, drugs, and a pre-fabricated drug matrix.

[0145] Item 21. A kit compatible with the loading door comprising a loading system of the device according to any one of Items 1 to 20, wherein the kit comprises reagents as required for performing an assay cycle for one patient sample. Item 22. The kit according to Item 21, comprising an environmental monitoring tag for tracking the transport and handling conditions, wherein the monitoring tag optionally comprises means for wireless communication, such as, for example, an RFID.

[0146] Item 23. The kit according to Item 21 or 22, comprising a unique identifier, such as, for example a barcode or RFID, that is manually or automatically read upon loading of the kit in order to ensure a correct assignment of the sample and to identify the content thereof.

[0147] Item 24. The kit according to any one of Items 21 to 23, comprising a UV-resistant cover for allowing a UV decontamination inside the loading system.

[0148] Item 25. The kit according to Item 24, wherein the kit comprises spacer elements and a stacking plate to keep contents in place and to separate contents from cover to prevent sticking in a frozen state.

[0149] Item 26. The kit according to Item 24 or 25, wherein the cover is locked to a base plate to ensure a secure transport prevent contamination, in particular in a frozen state.

[0150] Item 27. The kit according to Item 26, wherein the lock is mechanically unlocked upon insertion into the loading system and the cover is opened upon opening of an inner loading system door.

[0151] Item 28. A continuous and automated method for identifying anticancer drugs or drug combinations for patients that suffer from, or are being diagnosed for, a neoplastic disease or tumor, the method comprising a) providing a laboratory automation device according to any one of Items 1 to 20, b) providing at least one first sample, such as a patient-derived sample, comprising cancer cells to the laboratory automation device according to step a), and starting a first identification cycle at starting timepoint Tl, wherein the first identification cycle comprises the steps of i) optionally, dissociating the cancer cells of the sample of step b) a in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on the cells, in particular the dissociated cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drug and / or combination of drugs, iv) determining an anti-cancer effect of the drugs and / or combination of drugs of step iii) on said array of said 3D microtissues, and iv) identifying an anti cancer drug or drug combination based on the effect as determined in step iii) at timepoint T2, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified; c) providing at least one second sample, such as a patient-derived sample, comprising cancer cells to the laboratory automation device according to step a) through the loading system thereof, and starting a second identification cycle at starting timepoint T3, wherein timepoint T3 is after T1 and before T4, wherein the second identification cycle comprises the steps of i) optionally, dissociating the cancer cells of the sample of step b), in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on the cells, in particular the dissociated cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drug and / or combination of drugs, iv) determining an anti-cancer effect of the drug and / or combination thereof of step iii) on said array of said 3D microtissues, and iv) identifying an anticancer drug or drug combination based on the effect as determined in step iii) at timepoint T4, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified.

[0152] Item 29. The method according to Item 28, wherein the first sample, such as the patient- derived sample, comprising cancer cells is provided to the laboratory automation device according to step a) through the loading system thereof.

[0153] Item 30. The method according to Item 28 or 29, wherein the anticancer drug or drug combination is furthermore identified as patient-specific and / or personalized.

[0154] Item 31. The method according to any one of Items 28 to 30, furthermore comprising the step of storing the results of the identifying step on a suitable storage medium, such as a computer.

[0155] Item 32. The method according to any one of Items 28 to 31, wherein 2, 3, 4, 5, 6, or more independent identification cycles are performed, preferably at different starting timepoints Tx, wherein X is an integer corresponding to the beginning of the independent identification cycle as performed.

[0156] Item 33. The method according to any one of Items 28 to 32, further comprising a cycleindependent sample loading through a loading system.

[0157] Item 34. The method according to any one of Items 28 to 33, further comprising an automated prediction and pre-notification of a user to prevent waste overspills, consumables shortages or device interruptions.

[0158] Item 35. The method according to any one of Items 28 to 34, further comprising a selforganized prioritization of samples, critical process steps and timing constraints by the device, such as, for example cycle incubation times.

[0159] Item 36. The method according to any one of Items 28 to 35, further comprising the step of detection of contamination and an automatic elimination of contaminated samples, preferably further comprising an isolation and export of the contaminated samples.

[0160] Item 37. The method according to Item 36, wherein the detection comprises a machine learning-based recognition of contaminations, such as, for example, fungi and / or an image trend analysis-based recognition of contaminations, such as, for example, bacteria.

[0161] Item 38. The method according to any one of Items 28 to 37, further comprising an in- situ decontamination of materials as loaded through the loading system.

[0162] Item 39. The method according to any one of Items 28 to 38, further comprising the step of an automatic decontamination of the at least one device during idle times before, during or after performing the method.

[0163] Item 40. The method according to any one of Items 28 to 39, wherein dissociating said tissue sample comprises i) if required, dissecting said tissue sample into smaller pieces comprising cells, ii) treating said tissue sample with a solution comprising at least one enzyme capable of dissociating cells in said tissue sample, preferably at least one enzyme selected from a protease, a collagenase, trypsin, elastase, hyaluronidase, papain, chymotrypsin, deoxyribonuclease I, and neutral protease (dispase), producing a supernatant comprising dissociated cells, and ii) removing said supernatant comprises said dissociated cells and suitably collecting said cells, wherein steps (ii) and (iii) are repeated at least once, wherein preferably before step (ii) said tissue sample is sonicated with ultrasound, wherein the energy of said ultrasound is set at a suitable level to not destroy a substantial amount of said cells.

[0164] Item 41. The method according to any one of Items 28 to 40, wherein step b) comprises adding or removing stroma cells, stromal fibroblasts, endothelial cells and immune cells to said dissociated cells, and / or wherein in step b) for each 3D microtissue a predetermined number of cells is provided, such as, for example, between 500 and 10000 viable cells, and / or wherein in step b) said 3D microtissues are generated in at least one system selected from a hanging drop system, and b) a multiwell system, preferably comprising Ultra Low Adherence (ULA) wells.

[0165] Item 42. The method according to any one of Items 28 to 41, wherein the generation of said 3D microtissues does not require the use of a solubilized basement membrane preparation, like Matrigel® and / or wherein the generation of said 3D microtissues comprises self-assembly of said cells comprised in said dissociated cells, and / or wherein the generation of said 3D microtissues comprises a maturation time of about 6 hours to 7 days, preferably about 1 to 6 days, more preferably about 2 to 5 days, and / or wherein said 3D microtissues as generated have a size of 350 pm + / - 100 pm.

[0166] Item 43. The method according to any one of Items 28 to 42, wherein said contacting in step c) comprises a continuous exposure to said at least one drug and / or combination of drugs, and / or an exposure to and subsequent removal to said at least one drug and / or combination of drugs.

[0167] Item 44. The method according to any one of Items 28 to 43, wherein said determining of said effect in step d) is selected from size determination of said 3D microtissue, quantification of internal reporter gene expression in said 3D microtissue, determination of the intracellular ATP content in said 3D microtissue, and determination of pre-selected biomarkers in said 3D microtissue, wherein preferably said size determination of said 3D microtissue comprises at least one parameter selected from diameter, perimeter, volume, and area of optical cross section, and wherein preferably said size determination of said 3D microtissue comprises the use of an imaging device, and optionally further comprising the analysis of growth-kinetics.

[0168] Item 45. The method according to any one of Items 28 to 44, wherein said sample comprises cells, such as a patient-derived tissue sample selected from a sub-sample derived from a primary tissue sample, a primary tumor sample, and a metastasis sample, and wherein preferably said tissue sample has been obtained by a method comprising core biopsy, tumor resection, liquid biopsy and / or needle aspiration, and / or wherein said tissue sample and / or the dissociated cells are frozen and re-thawed prior to the generation of said 3D microtissues.

[0169] Item 46. The method according to any one of Items 28 to 45, comprising providing a sample, e.g. providing a primary tissue sample, obtaining a subsample in addition to the sample, such as the patient-derived sample and subjecting said subsample to at least one of molecular profiling, histological analysis, and histochemical analysis.

[0170] Item 47. Use of the device according to any one of Items 1 to 20 or the kit according to any one of Items 21 to 27 in an automated and continuous method for identifying an anticancer drug or anti-cancer drug according to the method according to any one of Items 28 to 46.

[0171] The invention will now be described further in the following examples with reference to the accompanying figures, nevertheless, without being limited thereto. For the purposes of the present invention, all references as cited are incorporated by reference in their entireties.

[0172] In the accompanying figures,

[0173] Figure 1 shows an illustrative drawing of the loading door and lock system as well as a kit according to the invention.

[0174] Figure 2 shows an illustrative drawing of the kit, box and container according to the invention.

[0175] Figure 3 shows another illustrative drawing of the loading door and lock system as well as a kit according to the invention.

[0176] Figure 4 shows another illustrative drawing of the kit, box and container according to the invention.

Claims

Claims1. A laboratory automation device for an automated and continuous method for identifying an anti-cancer drug or anti-cancer drug combination, the device comprising i) at least one workspace area enclosed by at least one housing; ii) at least one automated handling arm for handling samples, drug samples, and / or cell culture materials, iii) optionally, at least one tissue sample dissociation unit for dissociating patient- derived cancer cell comprising tissue samples in order to obtain dissociated cancer cells, iv) at least one unit for producing at least one array of 3D microtissues, such as microtumors, based on said cancer cells, in particular the dissociated cancer cells of step ii), v) at least one drug testing unit for contacting the at least one array of said 3D microtissues with at least one potential anti-cancer drug and / or combination of drugs, vi) at least one first analysis unit for determining an effect of said drug and / or combinations thereof on the at least one array of said 3D microtissues as produced in step iv), vii) at least one second analysis unit for identifying an anti-cancer drug or anticancer drug combination based on the effect as determined in step vi), and optionally further comprising a unit for selecting said anti-cancer drugs or anticancer drug combinations as identified, and viii) the housing comprising at least one loading door comprising a loading system suitable for a sterile loading and unloading of patient samples.

2. The device according to claim 1, further comprising a second loading door comprising a loading and unloading system for a sterile feeding or removing of cell culture consumable materials.

3. The device according to claim 1 or 2, wherein said loading system is adapted tospecifically fit to a transport box or container, wherein preferably said transport box or container comprises at least one door to be opened and closed inside the system.

4. The device according to any one of claims 1 to 3, wherein the workspace area comprises 1, 2, 3, 4 or 5 of the units in one housing, preferably, wherein preferably all units except the at least one second analysis unit are enclosed by one housing.

5. The device according to any one of claims 1 to 4, wherein any separately housed units are interconnected with suitable doors and / or locks providing an uninterrupted and sterile loading and unloading of patient samples and / or materials between the units.

6. The device according to any one of claims 1 to 5, wherein said tissue sample dissociation unit and said unit for producing an array of 3D microtissues are positioned in the same housing, and / or wherein said drug testing unit and said first analysis unit are positioned in the same housing, and preferably wherein said two housings are interconnected to form a discrete device and / or wherein said device is, at least in part, arranged vertically.

7. The device according to any one of claims 1 to 6, wherein the loading system further comprises means for sterilizing the materials, such as, for example, a lock system for sterilizing by UV, and / or wherein said lock system further comprises means for thawing or cooling / freezing the materials to be loaded or unloaded.

8. The device according to any one of claims 1 to 7, further comprising implemented computer program means for automatically operating, monitoring, organizing and prioritizing all samples and assay steps, in order to ensure an uninterrupted and most efficient execution of all device activities.

9. The device according to claim 8, wherein the means for organizing further provide a self-organized prioritization of critical process steps and timing constraints, such as, for example, incubation times, and / or means for predicting and pre-notification of a user to prevent waste overspills, consumables shortages and / or deviceinterruptions.

10. The device according to any one of claims 1 to 9, further comprising means to automatically detect and remove contaminations, in particular comprising means for machine learning-based recognition of contaminations, such as fungi, and / or means for image trend analysis-based recognition of contamination, such as bacteria.

11. The device according to claim 10, wherein the means provide an automatic decontamination of the device during idle times.

12. The device according to any one of claims 1 to 11, further comprising computer program means to ensure a correct assignment of a kit, box or container as used to a patient sample, and / or a logging of all device activities.

13. The device according to any one of claims 1 to 12, wherein the at least one automated handling arm for handling samples, drug samples, and / or cell culture materials is a robotic arm comprising suitable means for gripping samples, drug samples, and / or cell culture materials.

14. The device according to any one of claims 1 to 13, wherein said tissue sample dissociation unit comprises at least one of i) a pipetting unit, ii) an enzyme reservoir, iii) a reservoir for cell culture media, iv) a reservoir for washing solutions, v) optionally, an ultrasonic device, and vi) a centrifuge unit, and / or wherein said unit for producing an array of 3D microtissues, such as microtumors, based on said dissociated cells comprises at least one of i) a pipetting unit, ii) a cell counting unit, and, iii) a handler for microtiter plates, and / or wherein said drug testing unit comprises at least one of i) a handler for microtiter plates, ii) a pipetting unit, iii) a reservoir for cell culture media, iv) an array of reservoirs comprising at least two different drugs or combinations thereof, and iv) an incubator unit, and / or wherein said first and / or second analysis unit comprises i) a handler for microtiter plates, and / or ii) an imaging system comprising a microscope and a camera, and optionally an HR scanner.

15. The device according to claim 14, wherein said tissue sample dissociation unit and said unit for the production of an array of 3D microtissues share the same pipetting unit and / or wherein said drug testing unit and said first analysis unit share the same handler for microtiter plates.

16. The device according to any one of claims 1 to 15, wherein the loading door comprising the loading system comprises an inner and outer door, wherein the outer door can be locked, in particular automatically, in order to prevent user manipulation and contamination.

17. The device according to any one of claims 1 to 16, wherein the loading system comprises means selected from UV decontamination means or a HEPA filtered air exchange for decontaminating the system after a loading or unloading, decontaminating a container cover, such as a kit cover, after loading, and / or decontaminating a contaminated test plate, if necessary.

18. The device according to any one of claims 1 to 17, wherein the loading system comprises means for unlocking the cover and opening of a kit upon insertion thereof.

19. The device according to any one of claims 1 to 18, wherein the loading system comprises a tracking system, such as, for example an RFID or barcode reader.

20. The device according to any one of claims 1 to 19, wherein the patient samples or cell culture consumable materials are selected from the group consisting of microtumors, biopsy material, digestion enzymes, culture medium, cells, cancer cells, supporting cells, control cells, cancer-type specific maintenance medium, drugs, and a pre-fabricated drug matrix.

21. A kit compatible with the loading door comprising a loading system of the device according to any one of claims 1 to 20, wherein the kit comprises all reagents as required for performing a complete assay cycle for one patient sample.

22. The kit according to claim 21, comprising an environmental monitoring tag for tracking the transport and handling conditions, wherein the monitoring tag optionally comprises means for wireless communication, such as, for example, an RFID.

23. The kit according to claim 21 or 22, comprising a unique identifier, such as, for example a barcode or RFID, that is manually or automatically read upon loading of the kit in order to ensure a correct assignment of the sample and to identify the content thereof.

24. The kit according to any one of claims 21 to 23, comprising a UV-resistant cover for allowing a UV decontamination inside the loading system.

25. The kit according to claim 24, wherein the cover comprises a stacking plate to keep contents in place and to separate contents from cover to prevent sticking in a frozen state.

26. The kit according to claim 24 or 25, wherein the cover is locked to a base plate to ensure a secure transport prevent contamination, in particular in a frozen state.

27. The kit according to claim 26, wherein the lock is mechanically unlocked upon insertion into the loading system and the cover is opened upon opening of an inner loading system door.

28. A continuous and automated method for identifying anticancer drugs or drug combinations for patients that suffer from, or are being diagnosed for, a neoplastic disease or tumor, the method comprising a) providing a laboratory automation device according to any one of claims 1 to 20, b) providing at least one first sample, such as a patient-derived sample, comprising cancer cells to the laboratory automation device according to step a), and starting a first identification cycle at starting timepoint Tl, wherein the first identification cycle comprises the steps ofi) optionally, dissociating the cancer cells of the sample of step b) a in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on said dissociated cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drug and / or combination of drugs, iv) determining an anti-cancer effect of the drug and / or combination of drugs of step iii) on said array of said 3D microtissues, and iv) identifying an anticancer drug or drug combination based on the effect as determined in step iii) at timepoint T2, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified; c) providing at least one second sample, such as a patient-derived sample, comprising cancer cells to the laboratory automation device according to step a) through the loading system thereof, and starting a second identification cycle at starting timepoint T3, wherein timepoint T3 is after T1 and before T4, wherein the second identification cycle comprises the steps of i) optionally, dissociating the cancer cells of the sample of step b), in order to obtain dissociated cancer cells, ii) generating an array of 3D microtissues, such as microtumors, based on said dissociated cells of step i), iii) contacting said array of said 3D microtissues with at least one potential anticancer drug and / or combination of drugs, iv) determining an anti-cancer effect of the drug and / or combination of drugs of step iii) on said array of said 3D microtissues, and iv) identifying an anticancer drug or drug combination based on the effect as determined in step iii) at timepoint T4, and optionally, further comprising the step of selecting said patient-specific drug or drug combination as identified.

29. The method according to claim 28, wherein the first sample such as the patient- derived sample, comprising cancer cells is provided to the laboratory automation device according to step a) through the loading system thereof.

30. The method according to claim 28 or 29, wherein the anticancer drug or drugcombination is furthermore identified as patient-specific and / or personalized.

31. The method according to any one of claims 28 to 30, furthermore comprising the step of storing the results of the identifying step on a suitable storage medium, such as a computer.

32. The method according to any one of claims 28 to 31, wherein 2, 3, 4, 5, 6, or more independent identification cycles are performed, preferably at different starting timepoints Tx, wherein X is an integer corresponding to the beginning of the independent identification cycle as performed.

33. The method according to any one of claims 28 to 32, further comprising a cycleindependent sample loading through a loading system.

34. The method according to any one of claims 28 to 33, further comprising an automated prediction and pre-notification of a user to prevent waste overspills, consumables shortages or device interruptions.

35. The method according to any one of claims 28 to 34, further comprising a selforganized prioritization of samples, critical process steps and timing constraints by the device, such as, for example cycle incubation times.

36. The method according to any one of claims 28 to 35, further comprising the step of detection of contamination and an automatic elimination of contaminated samples, preferably further comprising an isolation and export of the contaminated samples.

37. The method according to claim 36, wherein the detection comprises a machine learning-based recognition of contaminations, such as, for example, fungi and / or an image trend analysis-based recognition of contaminations, such as, for example, bacteria.

38. The method according to any one of claims 28 to 37, further comprising an in-situ decontamination of kits as loaded through the loading system.

39. The method according to any one of claims 28 to 38, further comprising the step of an automatic decontamination of the at least one device during idle times before, during or after performing the method.

40. The method according to any one of claims 28 to 39, wherein dissociating said tissue sample comprises i) if required, dissecting said tissue sample into smaller pieces comprising cells, ii) treating said tissue sample with a solution comprising at least one enzyme capable of dissociating cells in said tissue sample, preferably at least one enzyme selected from a protease, a collagenase, trypsin, elastase, hyaluronidase, papain, chymotrypsin, deoxyribonuclease I, and neutral protease (dispase), producing a supernatant comprising dissociated cells, and ii) removing said supernatant comprises said dissociated cells and suitably collecting said cells, wherein steps (ii) and (iii) are repeated at least once, wherein preferably before step (ii) said tissue sample is sonicated with ultrasound, wherein the energy of said ultrasound is set at a suitable level to not destroy a substantial amount of said cells.

41. The method according to any one of claims 28 to 40, wherein step b) comprises adding or removing stroma cells, stromal fibroblasts, endothelial cells and immune cells to said dissociated cells, and / or wherein in step b) for each 3D microtissue a predetermined number of cells is provided, such as, for example, between 500 and 10000 viable cells, and / or wherein in step b) said 3D microtissues are generated in at least one system selected from a hanging drop system, and b) a multiwell system, preferably comprising Ultra Low Adherence (ULA) wells.

42. The method according to any one of claims 28 to 41, wherein the generation of said 3D microtissues does not require the use of a solubilized basement membrane preparation, like Matrigel® and / or wherein the generation of said 3D microtissues comprises self-assembly of said cells comprised in said dissociated cells, and / or wherein the generation of said 3D microtissues comprises a maturation time of about 6 hours to 7 days, preferably about 1 to 6 days, more preferably about 2 to 5days, and / or wherein said 3D microtissues as generated have a size of 350 gm + / - 100 gm.

43. The method according to any one of claims 28 to 42, wherein said contacting in step c) comprises a continuous exposure to said at least one drug and / or combination of drugs, and / or an exposure to and subsequent removal to said at least one drug and / or combination of drugs.

44. The method according to any one of claims 28 to 43, wherein said determining of said effect in step d) is selected from size determination of said 3D microtissue, quantification of internal reporter gene expression in said 3D microtissue, determination of the intracellular ATP content in said 3D microtissue, and determination of pre-selected biomarkers in said 3D microtissue, wherein preferably said size determination of said 3D microtissue comprises at least one parameter selected from diameter, perimeter, volume, and area of optical cross section, and wherein preferably said size determination of said 3D microtissue comprises the use of an imaging device, and optionally further comprising the analysis of growth-kinetics.

45. The method according to any one of claims 28 to 44, wherein said sample, such as the patient-derived tissue sample, is selected from cells or a sub-sample derived from a primary tissue sample, a primary tumor sample, and a metastasis sample, and wherein preferably said tissue sample has been obtained by a method comprising core biopsy, tumor resection, liquid biopsy and / or needle aspiration, and / or wherein said cell or tissue sample and / or the dissociated cells are frozen and re-thawed prior to the generation of said 3D microtissues.

46. The method according to any one of claims 28 to 45, comprising providing cells or a primary tissue sample, obtaining a subsample in addition to the patient-derived sample and subjecting said subsample to at least one of molecular profiling, histological analysis, and histochemical analysis.

47. Use of the device according to any one of claims 1 to 20 or the kit according to anyone of claims 21 to 27 in an automated and continuous method for identifying an anti-cancer drug or anti-cancer drug according to the method according to any one of claims 28 to 46.

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