METHOD FOR THE PROCUREMENT AND / OR ARRANGEMENT OF CELL CULTURES

Description

[0001] The invention relates to a method for producing and / or arranging cell cultures and / or cell agglomerations.

[0002] For the development of cell cultures, it is common practice to fill or apply living cells to so-called cell carriers. These cell carriers can be configured, for example, as arrays or hanging drop plates. Cell cultures in cell carriers can be used, for instance, for medical or pharmacological studies.

[0003] Filling or applying such cell carriers can be done, for example, using pipettes or pipette systems. However, this can subject the cells to high shear stress, which can reduce their survival rate. Depending on the design of the pipettes or pipette systems, filling the cell carriers with cells or applying the cells to the carriers can require many individual steps, thus increasing the overall process time. The remaining study time for the respective cell culture can therefore be reduced, as several cell divisions may have already occurred by the time the filling or application process is complete.

[0004] WO 2013 / 113883 discloses a method for producing a three-dimensional structure from a structural material containing living cells, such as endothelial cells, nerve cells, osteoblasts, chondrocytes in a liquid by printing or extrusion.

[0005] Against the background set out above, the object of the present invention was to provide a method for the production and / or arrangement of cell cultures and / or cell agglomerations that can be carried out with improved productivity and at the same time a reduced risk of cell damage. Likewise, the object was to provide an arrangement of a plurality of cell cultures and / or cell agglomerations.

[0006] With regard to the method, this problem has been solved by the subject matter of claim 1.

[0007] Advantageous configurations are the subject of the dependent claims and are explained below.

[0008] In a method according to the invention for the production and / or arrangement of cell cultures and / or cell agglomerations, a printing medium containing living cells is provided and the printing medium is printed through a printing screen and / or a printing stencil.

[0009] Cell cultures and / or cell agglomerations produced and / or arranged by a method according to the invention are particularly suitable for carrying out medical and / or pharmacological investigations. A high degree of productivity can be advantageously ensured by printing the printing medium through a printing screen and / or a printing stencil. In particular, the design of the printing screen and / or the printing stencil allows the printing of several defined print areas or print sections in just one printing step. The entire printing process can thus be completed in a very short time. The remaining examination period of the respective cell culture and / or cell agglomeration can therefore be extended.

[0010] At the same time, the printing screen or printing stencil can advantageously reduce the shear stresses on the cells in the printing medium. In particular, the openings of the printing screen and / or the printing stencil can be designed to be sufficiently large to reduce shear stresses.

[0011] This reduces the risk of cell damage in the printing medium during the printing or application process. It also improves the suitability of the cell culture and / or cell agglomeration for medical and / or pharmacological investigations.

[0012] According to the present invention, a printing screen can be equipped with a fabric or formed by a fabric and / or by a frame for the fabric. Such a fabric can be designed for pressing a printing paste through it. The fabric of a printing screen can be arranged in a frame or clamped in place. Furthermore, a fabric can be formed by metal and / or plastic threads. Such metal and / or plastic threads can be arranged twisted together or run individually.

[0013] To achieve the desired printing form, a printing screen can have permeable and impermeable areas. Permeable areas can be permeable to a printing medium, and impermeable areas can be impermeable to a printing medium. According to the present invention, permeable areas of a printing screen can form a passage and / or a cutout in which a fabric permeable to a printing medium is arranged. Impermeable areas can also be provided with a fabric, which, however, is designed to be impermeable to a printing medium or is covered to achieve impermeability.

[0014] Fine structures can be created using a printing screen, especially with a thickness between 15 µm and 500 µm. Printing with a printing screen can be carried out with relatively high accuracy, particularly in the edge areas of the printed product.

[0015] According to the present invention, a printing stencil can be equipped with openings and / or cutouts, in particular with through-openings and / or cutouts, which are formed in a plate. A plate with openings and / or cutouts can form a printing stencil according to the present invention. Accordingly, the openings and / or cutouts can form a passage for printing paste. Printing paste can be forced through such openings and / or cutouts during a printing process. A printing stencil can, in particular, be free of fabric. Specifically, the openings and / or cutouts of a printing stencil can be free of fabric.

[0016] Simultaneously, it is possible to arrange a form element within an opening and / or cutout of a printing stencil to define the shape of the opening and / or cutout according to the desired printing form. Such form elements can be held in the desired position within the respective opening and / or cutout by individual retaining threads or similar means.

[0017] Printing stencils are particularly suitable for printing layers with a thickness of 100 µm or more, especially 300 µm or more, and most preferably 500 µm or more. Printing using a printing stencil can be carried out at relatively high speeds, as larger layer thicknesses can be achieved.

[0018] According to an advantageous embodiment, a support structure can be provided for a plurality of separate cell cultures and / or cell agglomerations. Furthermore, the printing medium can be printed onto a support structure using a printing screen and / or a printing stencil. The support structure can, in particular, form a printing substrate onto which the printing medium is printed after passing through the printing screen and / or the printing stencil. Such a support structure can simplify the handling of the cell cultures and / or cell agglomerations, especially after printing is complete.

[0019] Furthermore, it is possible to print and provide a support structure together with, and / or alternately with, the printing medium containing the living cells. This ensures a high degree of manufacturing flexibility. The positioning and alignment of a finished support structure beneath a printing screen and / or stencil is therefore unnecessary. This reduces the preparation effort for printing.

[0020] It is also possible to print a support structure before printing the printing medium containing the living cells, and thus provide it in this way. The fully printed support structure can then be used as a printing substrate.

[0021] According to a further advantageous embodiment, single- or multi-layered cell agglomerations and / or single- or multi-layered cell cultures can be generated by the pressure of the printing medium. Printing a single-layered cell agglomeration and / or cell culture can be achieved with minimal effort and therefore at low cost. Multi-layered cell agglomerations and / or cell cultures enable particularly informative investigations, as the conditions of living organisms can be replicated especially well through multi-layered structures.

[0022] In a further preferred embodiment, a multilayered tissue can be produced simply and quickly by repeatedly printing a printing medium containing living cells and / or by varying the cells or the printing medium containing cells in a layering direction (z-direction). Such a multilayered tissue can be, for example, skin or other types of cellular tissue.

[0023] In a further preferred manner, the printing medium can be conveyed through the printing screen and / or the printing stencil and / or onto a support structure by at least one doctor blade operation and / or at least one doctor blade movement, in particular by a plurality of doctor blade operations and / or a plurality of doctor blade movements. Doctor blade operations or doctor blade movements allow for rapid and uniform printing of the printing medium through the printing screen or the printing stencil. In particular, highly reproducible results can be achieved in this way.

[0024] In a particularly preferred manner, the printing medium can be printed by at least one screen printing process, and in particular by a plurality of screen printing processes. The printing of the printing medium can be carried out, in particular, by screen printing and / or stencil printing, and especially preferably by 2D or 3D screen printing. This ensures a particularly high degree of productivity and accuracy. Relatively large volumes of a printing medium can be printed with minimal time and cost in screen printing. The arrangement and / or production of cell cultures and cell agglomerations can thus be carried out quickly and cost-effectively. At the same time, the use of a screen printing process, in particular a 2D or 3D screen printing process, ensures gentle processing of the printing medium containing living cells.The individual cells can be processed and / or printed using screen printing with only a low risk of damage from shear stress. A 3D screen printing system can be used, in particular, for printing the printing medium.

[0025] In a further preferred manner, substructures of cell cultures and / or cell agglomerations, or tissues produced therefrom, are printed with an accuracy of up to 100 cells, preferably up to 50 cells, particularly 20 cells, and more preferably up to 10 cells, particularly down to a single cell. Such printing or manufacturing accuracies can be achieved both in the layer build-up direction (z-direction) and / or in directions transverse to the layer build-up direction (x / y-directions). In particular, such accuracies can be achieved by means of 3D screen printing.

[0026] The screen printing process, or the multiple screen printing processes, can advantageously be carried out on, and in particular directly onto, a substrate. Screen printing on a substrate, for example a cell carrier, enables a high degree of reproducibility as well as ease of handling when removing the printed cells after completion of the printing process.

[0027] Furthermore, it can be advantageous to print the living cell-containing printing medium together with and / or alternately with the printing of the support structure using 2D or 3D screen printing. This allows for flexible design of the support structure as well as the entire printing process. The desired shape of the support structure can thus be produced according to specific requirements.

[0028] In a further preferred embodiment, a three-dimensional cell culture and / or cell agglomeration can be created by printing a substrate with the printing medium. Three-dimensional cell cultures and / or cell agglomerations can be produced in particular by repeated printing or by 3D screen printing.

[0029] Furthermore, it is possible that after printing a substrate with the printing medium, the respective cell culture and / or cell agglomeration develops into a cell culture and / or cell agglomeration, particularly a three-dimensional cell culture and / or cell agglomeration. Accordingly, a development period may be required between the completion of the printing process and the development of a cell culture or cell agglomeration. The properties and / or characteristics of the cell cultures or cell agglomerations can be influenced over the duration of this development period.

[0030] According to a further preferred embodiment, structural frameworks and / or scaffolds can be provided in the printing sections, i.e., the sections created by the pressure of the printing medium containing living cells. Likewise, structural frameworks and / or scaffolds can be printed together with the printing medium and / or be contained within it. Scaffolds and / or scaffolds are particularly advantageous for the production of three-dimensional structures. Cell cultures and / or cell agglomerations can thus be formed three-dimensionally with minimal effort and reproducibly.

[0031] When printing with a living cell-containing printing medium, cavities, recesses, linear or channel-shaped structures can be advantageously incorporated. Such shapes allow for improved functionalization of the cell cultures, cell agglomerations, and / or tissues produced with the printing medium. For example, scaffolds and / or blood vessels can be formed using such cavities, recesses, and / or structures.

[0032] In a particularly preferred manner, a combination of materials can be printed, especially to produce scaffolds and / or matrices with cells in a single printing process with minimal effort.

[0033] In a further preferred embodiment, the respective cell culture and / or cell agglomeration can develop into tissue after printing. Tissue development can be particularly advantageous for use in medical or pharmacological studies. Tissue developed in this way can also be used for medical treatment. Finally, tissue developed in this way can serve as food. In particular, the respective cell culture and / or cell agglomeration can develop into synthetic meat after printing, which may be suitable for consumption.

[0034] According to a further advantageous embodiment, the support structure can be designed for the arrangement and / or holding of a plurality of separate cell cultures and / or cell agglomerations. In particular, the support structure can have predefined arrangement and / or holding sections for separate cell cultures and / or cell agglomerations. This ensures a secure and defined arrangement of the printed cells or the cell cultures and / or cell agglomerations formed by the cells on the support structure.

[0035] Preferably, the arrangement and / or receiving sections can be provided in multiple rows and columns on the support structure. In particular, the number of rows can differ from the number of columns. The external shape of the support structure can be adapted to the number of rows and columns. More preferably, the support structure can have recesses for receiving the printing medium and / or the arrangement and / or receiving sections can be designed as recesses.

[0036] The support structure can preferably be configured as a microscope slide and / or cell culture plate and / or cell carrier. This makes the support structure particularly suitable for use and / or handling in laboratories or for standardized transport containers, transport devices and / or storage devices.

[0037] In a further preferred embodiment, the printing screen and / or the printing stencil can have a plurality of cutouts for the passage of the printing medium, in particular, cutouts formed separately from one another. This allows a defined passage of the printing medium through the printing screen and / or the printing stencil to be achieved. In particular, this allows the printing medium to be printed and / or applied to a substrate in a defined manner at different and / or spaced-apart positions.

[0038] In a further preferred configuration, the cutouts of the printing screen and / or the printing stencil can correspond to predefined arrangement and / or receiving sections of the support structure. This allows the printing medium to be printed selectively in and / or onto the arrangement and / or receiving sections of the support structure. In particular, such a design prevents accidental or unwanted printing of the support structure outside the arrangement and / or receiving sections. Simultaneously, this enables a high printing volume per print run.

[0039] According to a further preferred embodiment, at least one cutout and / or passage of the printing screen and / or the printing stencil for the passage of the printing medium can have a size and / or a diameter of at least 1 mm, in particular at least 2 mm, preferably at least 2 mm, preferably at least 3 mm, more preferably 4 mm, more preferably at least 5 mm.

[0040] According to a further preferred embodiment, at least one cutout and / or passage of the printing screen and / or the printing stencil for the passage of the printing medium can have a size and / or diameter of up to 100 mm, in particular up to 50 mm, preferably up to 40 mm, more preferably up to 30 mm, more preferably up to 25 mm, more preferably up to 20 mm, even more preferably up to 15 mm, more preferably up to 10 mm, more preferably up to 9 mm, more preferably up to 8 mm, and more preferably up to 5 mm.

[0041] According to a further preferred embodiment, at least one cutout and / or passage of the printing screen and / or the printing stencil for the passage of the printing medium can have a size and / or diameter of 1 mm to 100 mm, in particular from 2 mm to 50 mm, preferably from 2 mm to 40 mm, more preferably from 2 mm to 30 mm, more preferably from 3 mm to 30 mm, more preferably from 3 mm to 25 mm, more preferably from 3 mm to 20 mm, even more preferably from 4 mm to 20 mm, more preferably from 4 mm to 15 mm, more preferably from 4 mm to 10 mm, even more preferably from 4 mm to 9 mm, more preferably from 4 mm to 8 mm, even more preferably from 5 mm to 10 mm, more preferably from 5 mm to 8 mm.By using such a dimensioned cutout and / or opening in the printing screen and / or in the printing stencil, on the one hand only a low shear stress on the living cells in the printing medium and at the same time a relatively large printing volume per printing process can be ensured.

[0042] According to a preferred embodiment, a plurality of arrangement and / or receiving sections of the support structure can be printed and / or filled with the printing medium simultaneously and / or in a single printing operation and / or by a single squeegee operation. This can be accomplished with minimal effort. In particular, all arrangement and / or receiving sections of the support structure can be printed with the printing medium in a single printing operation. The sequential filling of individual receiving sections in numerous individual steps can thus be avoided.

[0043] It is still possible to print and / or fill different arrangement and / or receiving sections of the carrier structure in sequential order, according to their spacing and the respective squeegee speed. The individual arrangement and / or receiving sections of the carrier structure can thus be filled according to a single squeegee movement. Simultaneous filling of the arrangement and / or receiving sections is therefore unnecessary, thus increasing process flexibility.

[0044] According to a further preferred embodiment of the method according to the invention, the printing medium can be printed onto a support structure at isolated positions. Printing the printing medium onto a support structure allows for the particularly advantageous creation of independent and / or fluidically separated printing sections. Consequently, several independent cell cultures and / or cell agglomerations can be generated, which are available for a relatively large number of medical and / or pharmacological investigations.

[0045] It can also be advantageous to overlay the printing medium with a sterile gas medium, particularly sterile air, during, before, and / or after printing. This ensures compliance with sterility requirements for handling and / or processing live cells. Unwanted contamination of cell cultures and / or cell agglomerations can thus be avoided.

[0046] The printing medium can be, in particular, a nutrient medium and / or a nutrient-containing medium, especially a nutrient liquid. Such a printing medium or nutrient medium can, in particular, be designed as so-called bio-ink.

[0047] According to a further preferred embodiment, the printing medium can be configured as a printing paste and / or a low-viscosity or medium-viscosity nutrient solution and / or a liquid suspension. In purely gel-like media, cells and / or nutrients can only diffuse to a limited extent, which can lead to restricted nutrient availability and thus also a reduced cell lifetime. By configuring the printing medium as a relatively low-viscosity printing paste and / or a low-viscosity or medium-viscosity nutrient solution and / or a liquid suspension, a high level of nutrient supply to the living cells within the printing medium can be ensured.

[0048] It is also possible that the printing medium is gel-like and / or designed as a highly viscous nutrient medium, which can simplify processing or printing.

[0049] Furthermore, the printing medium can be designed as a sol-gel matrix and / or have a variable viscosity and / or its viscosity can be modified by a drying step and / or a temperature control step, in particular by a decrease or increase in temperature. Depending on the processing step performed, the viscosity can thus be altered. This ensures a high level of nutrient supply over relatively long periods, while simultaneously allowing the viscosity to be modified for processing or further processing steps.

[0050] The printing medium can also be shear-thinning and / or shear-thinning. Under relatively high shear loads, this can result in a temporarily lower viscosity, thereby reducing the shear stress. This further reduces the risk of cell damage.

[0051] In a further preferred embodiment, a drying and / or tempering step of the printed printing medium can be performed between successive printing steps. It is also possible to generate a sol-gel transition after printing the printing medium, particularly through a drying and / or tempering step, and / or to perform a further printing process after a sol-gel transition has been generated. The properties of the printing medium, or of the cell culture and / or cell agglomeration produced thereby, can thus be specifically influenced depending on the respective process stage.

[0052] In a further preferred embodiment, the printing medium can be liquid or have a low viscosity during printing, thus enabling simple printing. Following printing, or the printing of a layer, the respective layer can be solidified or the viscosity of the printed medium can be increased, for example, by tempering and / or by creating a sol-gel transition. The respective printed object, or the cell culture and / or cell agglomeration produced by printing, can thereby be suitably built up in a layer-building direction (z-direction).

[0053] Once the printed object is fully built up in height (z-direction), it can be placed in a suitable mold and / or stencil, or the mold and / or stencil can be placed on top of the printed object to enclose it. The printed object can thus be confined in the x / y direction. The printed medium, or the cell culture and / or cell agglomeration created by printing, can be returned to a more fluid state or a state with lower viscosity without becoming runny. This allows nutrients to circulate freely again within the cell culture and / or cell agglomeration created by printing. Tissue can then develop in an improved manner.

[0054] In a further preferred embodiment, the printing medium can contain cells capable of division and / or cells inducible to division. Furthermore, the living cells of the printing medium can be human, animal, and / or plant cells. In particular, the printing medium can contain all types of human, animal, and / or plant cells. Most preferably, the living cells can be all cells capable of division or inducible to division of the human or animal body or of plants. This results in a particular suitability for medical and pharmacological investigations.

[0055] In a further preferred manner, the printing medium may contain living cells from the group of primary cells, in particular all types of human, animal and / or plant primary cells.

[0056] Likewise, the printing medium may contain living cells from the group of cell lines, in particular all types of established human, animal and / or plant cell lines.

[0057] Primary cells can be, for example, organ cells, in particular organ cells of all organs, skin cells, fibroblasts, chondroblasts, osteoblasts, muscle cells, cardiac muscle cells, nerve cells, liver cells, islet cells, vascular cells, glandular tissue cells, tumor cells, in particular tumor cells of all tumor tissues, stem cells, in particular hematopoietic, mesenchymal and / or neuronal stem cells and / or induced pluripotent stem cells from tissues such as adipose tissue, skin and / or umbilical cord, and / or totipotent stem cells, in particular egg cells and / or embryonic cells, and / or pluripotent, multipotent, oligopotent and / or unipotent stem cells, blood cells and / or immune cells.

[0058] Cell lines may, for example, consist of immortalized cells of the cell types mentioned above and / or cells of the tissue types mentioned above.

[0059] It can be further advantageous if the printing medium contains different cells and / or cell types, in particular different types of cells and / or cell types mentioned above. This can result in a further improved suitability of the generated cell cultures and / or cell agglomerations for medical and / or pharmacological investigations.

[0060] Preferably, a tissue produced from the printing medium can contain different cells and / or cell types, in particular different types of the cells and / or cell types mentioned above. This can also result in a further improved suitability of the produced tissue for medical and / or pharmacological investigations.

[0061] It can be further advantageous if the printing medium contains at least one growth factor and / or at least one protein and / or extracellular matrix protein, in particular a growth factor and / or protein from the cytokine group, especially as growth regulators, interferons and / or interleukins, membrane components, laminins, collagens, especially collagen type 4, proteoglycans, entactins, nidogens, cell adherence factors, especially fibronectin and / or vitronectin, growth factors, especially of the EGF family, the TGF family, PDGF, VEGF, somatomedins, especially IGF, NGF, PTGF, and / or protective factors, especially tissue-specific plasminogen activators, serum albumins and / or CMC. In a particularly preferred manner, the printing medium may contain at least one or more of these substances from the aforementioned groups.This may result in a further improved suitability of the cell cultures, cell agglomerations and / or tissues produced using the printing medium for carrying out medical and / or pharmacological investigations.

[0062] In a further preferred embodiment, the printing medium can contain cells with a diameter or cross-sectional length of 5-50 µm, 5-40 µm, 10-50 µm, 10-40 µm, 10-30 µm, 20-40 µm, 25-40 µm, or 25-30 µm. Furthermore, a cell culture can contain 5 to 1000 cells, in particular 10 to 1000 cells, 20 to 500 cells, 50 to 500 cells, 100 to 500 cells, 200 to 500 cells, or 300 to 500 cells. This allows for particularly informative investigations using the cell cultures and / or cell agglomerations generated by the printing medium.

[0063] Another independent aspect of the present invention relates to a method for generating and / or arranging cell cultures and / or cell agglomerations in which a support structure is provided for a plurality of separate cell cultures and a printing medium containing living cells is printed onto the support structure by means of screen printing.

[0064] A further independent aspect of the present invention relates to an arrangement of a plurality of cell cultures and / or cell agglomerations printed onto a support structure by a method described above.

[0065] A further independent aspect of the present invention relates to an arrangement of a plurality of tissues and / or tissue sections which are produced at least by printing a printing medium onto a support structure according to a method described above and subsequent and / or prior cell cultivation.

[0066] According to a particularly preferred embodiment, the cell cultures and / or cell agglomerations and / or tissues can be identical with respect to the metabolic state of the cells, cell age, and / or cell number and / or cell number dimension. It is also possible that the cell cultures and / or cell agglomerations and / or tissues exhibit deviations with respect to the metabolic state of the cells, cell age, and / or cell number of less than 20%, in particular less than 10%, less than 5%, less than 3%, less than 2%, or less than 1%. Such an arrangement allows the cell cultures and / or cell agglomerations and / or tissues to be used for a variety of investigations in which the investigation parameters can be appropriately varied between the individual cell cultures and / or cell agglomerations and / or tissues. Such series of investigations can be particularly informative.

[0067] In the present context, tissue can be understood to mean, in particular, cellular tissue.

[0068] Further embodiments of the present invention result from combinations of the features disclosed in the claims, the description, and the figures. The present invention is explained below with reference to exemplary embodiments and accompanying drawings.

[0069] They each show schematically: Fig. 1 shows a support structure for cell cultures and / or cell agglomerations and a pressure sieve or pressure stencil arranged above it in perspective view; Fig. 2 shows an arrangement of a plurality of cell cultures and / or cell agglomerations on a support structure according to Fig. 1 in perspective view.

[0070] In Fig. 1 A support structure 10 for cell cultures and / or cell agglomerations is shown in perspective. The support structure 10 can be, in particular, a cell culture plate, a microscope slide, or a so-called array for cell cultures and / or cell agglomerations. The support structure 10 can have a plurality of mounting sections 12.

[0071] The receiving sections 12 can be configured to receive cell cultures and / or cell agglomerations, which are not shown in detail here. In particular, a pressure medium containing living cells, which is not shown in detail here, can be conveyed into the receiving sections 12 by pressure, as explained in more detail below. Once such a pressure medium is arranged in the receiving sections 12, it can develop into cell cultures and / or cell agglomerations. Likewise, a pressure medium already consisting of and / or containing cell cultures and / or cell agglomerations can be arranged in the receiving sections 12 by pressure.

[0072] According to the exemplary design in Fig. 1 The receiving sections 12 can be formed by depressions, in particular by bowl-shaped and / or hemispherical and / or cylindrical and / or rectangular depressions. The in Fig. 1 The exemplary support structure 10 has a total of 24 receiving sections 12. The receiving sections 12 can, by way of example, be arranged in a total of 4 rows and 6 lines within the support structure 10. Of course, support structures 10 with a different number of receiving sections 12 or with different arrangements of receiving sections 12 may also be suitable.

[0073] In the Fig. 1 Furthermore, a printing stencil 14 is shown in a schematic perspective view. The printing stencil 14 can be formed by a plate 16 which has a plurality of cutouts 18 for the passage of a printing medium. The cutouts 18 are, in particular, formed separately from one another.

[0074] Instead of the printing stencil 14, a printing screen (not shown in detail here) equipped with a mesh can also be provided. Within such a mesh, areas permeable to printing paste and bordered by impermeable areas can be provided. Permeable areas are also referred to as cutouts in this document, although they may contain a mesh.

[0075] The cutouts 18 of the printing stencil 14 or of a printing screen not shown here can, for example, have a diameter of at least 1 mm, in particular at least 2 mm, preferably at least 3 mm, more preferably at least 4 mm, more preferably at least 5 mm.

[0076] The cutouts 18 of the printing stencil 14 or of a printing screen (not shown here) can have a diameter of 1 mm to 100 mm, particularly 2 mm to 50 mm, preferably 2 mm to 40 mm, more preferably 2 mm to 30 mm, more preferably 3 mm to 30 mm, more preferably 3 mm to 25 mm, more preferably 3 mm to 20 mm, more preferably 4 mm to 20 mm, more preferably 4 mm to 15 mm, more preferably 4 mm to 10 mm, more preferably 4 mm to 9 mm, more preferably 4 mm to 8 mm, more preferably 5 mm to 10 mm, more preferably 5 mm to 8 mm. The cutouts 18 of the printing stencil 14 can have different diameters or identical diameters.

[0077] Furthermore, the cutouts 18 of the printing template 14 or of a printing screen not shown here can correspond in a particularly preferred manner to the predefined receiving sections 12 of the support structure 10.

[0078] According to the present invention, a printing medium containing living cells is printed through a printing screen and / or through the printing stencil 14.

[0079] For printing the printing medium, the printing screen and / or the printing stencil 14 can be arranged above the support structure 10, as shown in Fig. 1 shown. This further in Fig. 1 The schematically shown squeegee 20 can be used to print a printing medium through the printing screen and / or the printing stencil 14. The printing medium can be printed through the printing screen and / or the printing stencil 14 by at least one squeegee pass and / or at least one squeegee movement, in particular by a plurality of squeegee passes and / or a plurality of squeegee movements of the squeegee 20. This allows the printing medium to be conveyed onto the support structure 10 with particular reliability.

[0080] The printing of the printing medium using the printing screen and / or the printing stencil 14 and / or the printing squeegee 20 can be carried out in particular using the screen printing process. The screen printing process can be, in particular, a 2D screen printing process or a 3D screen printing process.

[0081] Furthermore, the support structure 10 can be printed together with and / or alternately with the printing medium containing living cells. In particular, the printing of the printing medium containing living cells can be carried out together with and / or alternately with the printing of the support structure 10 using 3D screen printing. Alternatively, the support structure 10 can be printed before the printing medium containing living cells, especially using 3D screen printing. Likewise, the support structure 10 can be manufactured using other methods and provided before the printing medium containing living cells.

[0082] In Fig. 2 An arrangement 11 of a plurality of cell cultures 22 and / or cell agglomerations 24 on a support structure 10 is shown in perspective. Each of the cell cultures 22 and / or cell agglomerations 24 is arranged in one of the receiving sections 12. Single- or multi-layered cell cultures 22 and / or single- or multi-layered cell agglomerations 24 can be generated by the pressure of the printing medium, which is not shown in detail here. Furthermore, a three-dimensional cell culture 22 and / or cell agglomeration 24 can be generated by printing the support structure 10 with the printing medium. After printing the support structure 10 with the printing medium, the respective cell culture 22 and / or cell agglomeration 24 can develop into a three-dimensional cell culture 22 and / or cell agglomeration 24.

[0083] Furthermore, it is possible that structural frameworks and / or scaffolds are provided in printing sections 26 or recording sections 12, which are also not shown in detail here. Such structural frameworks and / or scaffolds may be printed together with the printing medium and / or be contained within it.

[0084] In a particularly preferred manner, a plurality of receiving sections 12 of the carrier structure 10 can be printed and / or filled with the printing medium simultaneously and / or in a single printing operation and / or by a single squeegee operation. Likewise, different receiving sections 12 of the carrier structure 10 can be printed and / or filled sequentially according to their spacing from one another and according to the respective squeegee speed.

[0085] By printing the carrier structure 10 with the printing medium, independent and / or fluidically separated printing sections 26 can be created. In particular, one printing section 26 can be created for each receiving section 12. A printing section 26 can form a cell culture 22 and / or a cell agglomeration 24, or develop into a cell culture 22 and / or a cell agglomeration 24. Furthermore, the cell cultures 22 and / or the cell agglomerations 24 can develop into tissue after printing.

[0086] At the in Fig. 2 In the arrangement 11 shown, the individual cell cultures 22 and / or cell agglomerations 24 can be identical with respect to the metabolic state of the cells, cell age, and / or cell number and / or cell number dimension. Likewise, it is possible that in the arrangement 11 according to Fig. 2Deviations in the metabolic state of the cells, cell age and / or cell number are less than 20%, in particular less than 10%, less than 5%, less than 3%, less than 2% or less than 1%.

[0087] The printing medium and / or the cell cultures 22 and / or cell agglomerations 24 produced therewith can contain cells capable of division and / or cells inducible to division. The living cells of the printing medium and / or the cell cultures 22 and / or cell agglomerations 24 can be human, animal, and / or plant cells. In particular, the printing medium and / or the cell cultures 22 and / or cell agglomerations 24 can contain all types of human, animal, and / or plant cells. Preferably, the living cells of the printing medium and / or the cell cultures 22 and / or cell agglomerations 24 can be all cells capable of division or inducible to division of the human or animal body or of plants.

[0088] Furthermore, the printing medium or a cell culture 22 and / or cell agglomeration 24 generated therefrom may contain living cells from the group of primary cells, in particular all types of human, animal, and / or plant primary cells. Likewise, the printing medium or a cell culture 22 and / or cell agglomeration 24 generated therefrom may contain living cells from the group of cell lines, in particular all types of established human, animal, and / or plant cell lines. Finally, the printing medium or a cell culture 22 and / or cell agglomeration 24 generated therefrom may contain different cells and / or cell types.

[0089] Furthermore, the printing medium or a cell culture 22 and / or cell agglomeration 24 generated therefrom may contain at least one growth factor and / or at least one protein and / or extracellular matrix protein, in particular a growth factor and / or protein from the group of cytokines, especially as growth regulators, interferons and / or interleukins, membrane components, laminins, collagens, especially collagen type 4, proteoglycans, entactins, nidogens, cell adherence factors, especially fibronectin and / or vitronectin, growth factors, especially of the EGF family, the TGF family, PDGF, VEGF, somatomedins, especially IGF, NGF, PTGF, and / or protective factors, especially tissue-specific plasminogen activators, serum albumins and / or CMC. Likewise, the printing medium or a cell culture 22 and / or cell agglomeration 24 generated therefrom may contain at least one or more of these substances from the aforementioned groups.

[0090] The cell cultures 22 and / or cell agglomerations 24 produced according to the above-described methods are particularly suitable for carrying out medical and / or pharmacological investigations.

Claims

1. Method for producing and / or arranging cell cultures (22) and / or cell agglomerations (24), in particular for carrying out medical and / or pharmacological examinations, in which a printing medium containing living cells is provided and in which the printing medium is printed through a printing screen and / or a printing stencil (14).

2. Method according to claim 1, characterized in that the printing medium is printed through a printing screen and / or a printing stencil (14) onto a carrier structure (10) and / or that a carrier structure (10) is provided for a plurality of separate cell cultures (22) and / or cell agglomerations (24) and / or that a carrier structure (10) is printed together with and / or alternately with the printing medium containing living cells and / or that a carrier structure (10) is printed before the printing medium containing living cells is printed.

3. Method according to claim 1 or 2, characterized in that the printing medium is transported through the printing screen and / or the printing stencil (14) and / or onto a carrier structure (10) by at least one squeegee operation and / or at least one printing squeegee movement, in particular by a plurality of squeegee operations and / or a plurality of printing squeegee movements.

4. Method according to one of the preceding claims, characterized in that the printing medium is printed by at least one screen printing operation, in particular by a plurality of screen printing operations, in particular onto a carrier structure (10), and / or that the printing of the printing medium is carried out using the screen printing process, in particular the 2D screen printing process or 3D screen printing process, and / or that the printing of the printing medium containing living cells is carried out together and / or alternately with the printing of the carrier structure (10) using the screen printing process and / or that single-layer or multi-layer cell cultures (22) and / or single-layer or multi-layer cell agglomerations (24) are produced by printing the printing medium.

5. Method according to one of the preceding claims, characterized in that by printing the printing medium onto a carrier structure (10) a three-dimensional cell culture (22) and / or cell agglomeration (24) is produced and / or that after printing the printing medium onto a carrier structure (10), the respective cell culture (22) and / or cell agglomeration (24) develops into a three-dimensional cell culture (22) and / or cell agglomeration (24) and / or that structural scaffolds and / or scaffolds are provided in printing sections (26) and / or that structural scaffolds and / or scaffolds are printed together with the printing medium and / or are contained therein and / or that the cell cultures (22) and / or the cell agglomerations (24) develop into tissue after printing.

6. Method according to one of claims 2 to 5, characterized in that the carrier structure (10) is designed for the arrangement and / or reception of a plurality of separate cell cultures (22) and / or cell agglomerations (24) and / or that the carrier structure (10) has predefined arrangement and / or reception sections (12) for separate cell cultures (22) and / or cell agglomerations (24) and / or that the arrangement and / or receiving sections (12) are provided in several rows and columns on the carrier structure (10) and / or that the carrier structure (10) is designed as a microscope slide and / or cell culture plate and / or has recesses for receiving the printing medium.

7. Method according to one of the preceding claims, characterized in that the printing screen and / or the printing stencil (14) has a plurality of cut-outs (18) for the passage of the printing medium, in particular cut-outs (18) formed separately from one another, wherein the cut-outs (18) of the printing screen and / or the printing stencil (14) preferably correspond to predefined arrangement and / or receiving sections (12) of the carrier structure (10).

8. Method according to one of the preceding claims, characterized in that at least one cut-out (18) of the printing screen and / or the printing stencil (14) for the passage of the printing medium has a size and / or a diameter of 1 mm to 100 mm, in particular from 2 mm to 50 mm, preferably from 2 mm to 40 mm, preferably from 2 mm to 30 mm, more preferably from 3 mm to 30 mm, more preferably from 3 mm to 25 mm, more preferably from 3 mm to 20 mm, even more preferably from 4 mm to 20 mm, more preferably from 4 mm to 15 mm, more preferably from 4 mm to 10 mm, even more preferably from 4 mm to 9 mm, more preferably from 4 mm to 8 mm, even more preferably from 5 mm to 10 mm, more preferably from 5 mm to 8 mm.

9. Method according to one of claims 2 to 8, characterized in that a plurality of arrangement and / or receiving sections (12) of the carrier structure (10) are printed and / or filled with the printing medium simultaneously and / or in a printing process and / or by a squeegee process, and / or that the printing medium is printed onto a carrier structure (10) at individual positions and / or that printing a carrier structure (10) with the printing medium produces printing sections (26) that are independent of one another and / or separated in terms of fluid technology.

10. Method according to one of the preceding claims, characterized in that during printing, before printing and / or after printing, the printing medium is overlaid with a sterile gas medium, in particular with sterile air.

11. Method according to one of the preceding claims, characterized in that the printing medium is formed as a printing paste and / or low-viscosity or medium-viscosity nutrient fluid and / or liquid suspension and / or solgel matrix and / or that the printing medium has a variable viscosity and / or that the viscosity of the printing medium can be changed by a drying step and / or temperature control step and / or that the printing medium is structurally viscous.

12. Method according to one of the preceding claims, characterized in that a drying and / or temperature control step of the printed printing medium takes place between successive printing steps and / or that a sol-gel transition is produced after printing the printing medium, in particular by a drying and / or temperature control step, and / or that a further printing process is carried out after a sol-gel transition has been produced.

13. Method for producing and / or arranging cell cultures (22) and / or cell agglomerations (24), in particular according to one of the preceding claims, in which a carrier structure (10) is provided for a plurality of separate cell cultures (22) and / or cell agglomerations (24) and a printing medium containing living cells is printed onto the carrier structure (10) by means of screen printing.