CELL CULTURE PLATE AND USES THEREOF

D E S C R I P T I O N

Technical field

[0001] The present disclosure relates to the field of cell plates, cell culturing, and in particular, it relates to dividable surfaces for cell culturing and cell passaging.

Background

[0002] Cell culture is a complex process by which cells are grown, outside of their natural environment, under controlled conditions. In practice, the term "cell culture" refers to the culturing of cells derived from multi-cellular eukaryotes, especially animal cells, but also cultures of plants, fungi, insects and microbes, including viruses, bacteria and protists.

[0003] Culture conditions vary widely for each cell type, and variation of conditions for a particular cell type can result in different phenotypes. The most commonly varied factor in culture systems are cell growth medium, cell density, temperature and gas mixture.

[0004] The growth of animal cells in vitro is a demanding task as these cells require many nutrients and typically grow only when attached to specially coated surfaces. Despite these difficulties, various types of animal cells, including both undifferentiated and differentiated ones, can be cultured successfully.

[0005] Plating density (number of cells per volume of culture medium) plays a critical role for some cell types. Cells can be grown either in suspension or as adherent cultures. Some cells naturally live in suspension, without being attached to a surface, such as cells that exist in the bloodstream. There are also cell lines that have been modified to be able to survive in suspension cultures so they can be grown to a higher density than adherent conditions would allow.

[0006] Adherent cells require a surface, such as tissue culture plastic or microcarrier, which may be coated with extracellular matrix components, such as collagen and laminin, to increase adhesion properties and provide other signals needed for growth and differentiation. Most cells derived from solid tissues are adherent.

[0007] Another type of adherent culture is organotypic cultures, which involve growing cells in a three- dimensional (3D) environment as opposed to two-dimensional (2D) culture dishes. This 3D culture system is biochemically and physiologically more similar to in vivo tissue, but is technically challenging to maintain because of many factors, e.g. diffusion, among others. [0008] To grow different types of cells, in particular animal cells, is therefore highly demanding. This demand has to be combined with growing cells in conditions of reproducibility and wherein it is possible to sample replicates from the same cell culture for each and different analytical procedure and to sample at different time-points for determining time-related descriptors. Thus, in current state-of-the art, cell cultures are performed in well plates. These well plates are usually composed of 6, 12, 24 or 96 physically separated wells, in which medium and cells are seeded, well by well. Wells are then sampled for estimation of variation of phenotype in the cell culture and/or sampled over time to estimate values such as growth rate or metabolic consumption. However, the variation of phenotype between wells observed in the referred well plates renders the comparison of experiments very difficult due to the high associated error to each measurement of a replicated sample.

[0009] These facts are disclosed in order to illustrate the technical problem addressed by the present disclosure.

General description

[0010] The present disclosure may be used for any type of cell plate application and simultaneously it may be used for assay compartmentalization, probe multiple gene sub-set assays, wash Western blot strips, immunoassays, live/dead imaging assays, cell sampling assays for PC or Western blots, among others.

[0011] The cell plates already known for culturing cells, in particular for culturing animal cells, present several drawbacks, namely low reproducibility between wells, high number of handling steps, and high time-consumption during: well-plate coating, cell seeding, media exchanges and cell passaging.

[0012] The low reproducibility, or lack of reproducibility, between wells has the inconvenient of rendering high errors associated with the analytical measurement of a replicated sample, and of rendering high errors to the estimation of time-related descriptors of cell cultures such as growth curves, metabolic consumptions and profiles of labelled intracellular metabolites. These errors prevent an effective statistical comparison between experiments, either by attempting to prove differences or similarities.

[0013] The low reproducibility, or lack of reproducibility, between different wells of a cell well plate is a consequence of several factors, namely different evaporation rates of water in medium/buffer from different wells that occur in incubators, which leads to some wells having culture medium more concentrated than others; different concentration of cells between wells due to the seeding technique of pipetting well by well; for cell cultures involving changes in culture medium, there is a culture medium volume variation between wells, due to error in the pipetting volume of fresh culture medium discharged for each well. [0014] Thus, one of the objectives of this disclosure is to increase the reproducibility between different wells of a cell well plate.

[0015] The high number of handling steps has the inconvenient of increasing the overall propagated error of the operation as each step is associated with an error, the inconvenient of increasing the probability of contamination of wells or even of the whole well plate, and the inconvenient of consuming more labour time.

[0016] Therefore, one of the objectives of this disclosure is to reduce the number of handling steps.

[0017] The above-mentioned factors lead to different wells having different conditions, for example different ratios of culture medium volume/cell number, different ratios of cell number/cm ² of plate, resulting in differences in phenotype between wells at the end of culture. Thus, a need exists for a new cell culture tool that facilitates and enhances data reproducibility and reduces the number of handling steps.

[0018] The present disclosure provides a cell plate that simultaneously is responsible for increasing the reproducibility between different wells of a cell well plate and reduces the number of handling steps.

[0019] The present disclosure relates to a novel cell culture plate. With a single pipetting, one may distribute a cell culture medium and inoculate said medium with the desired cells. After inoculating the cell culture medium with the desired cell culture, one may lay one or more partitions on the cell plate, thereby creating physically separated and sealed compartments. Once two or more compartments are created, one can proceed to the sampling of the culture medium and/or sampling of cell cultures, in particular animal cell cultures.

[0020] In an embodiment, the compartments may be created by the positioning of one or more separators/partitions. The number of partitions may be made variable in number and size, and throughout the culture time.

[0021] This disclosure provides the technical advantage that all samplings have the same initial cell seeding and share the same culture medium, therefore reducing or avoiding the lack of reproducibility that characterise the already existing cell plates.

[0022] This disclosure may be used for animal cells either when the animal cell is the final product or when it is used as a mean for virus/vaccine/VLP or protein production. This disclosure may also be used for other type of cells such as fungi cells, bacteria cells or any other type of cells.

[0023] This disclosure may be used for adherent cells, cells in suspension, cells in microcarriers, 3D aggregates, encapsulated cells, spheres, spheroids, organoids or tumoroids, either when the cells are the final product or when it is used as a mean for virus/vaccine/VLP or protein production. [0024] For many applications, an extracellular matrix (ECM) coating of the tissue culture plastic and/or cell plates may improve cell attachment, differentiation, signalling, and increase adhesion properties and provide other signals needed for growth and differentiation.

[0025] In an embodiment, said coating may be a coating of collagen, poly-L-Lysine, poly-D-Lysine, fibronectin, vitronectin, hyaluronan, fibroin silk, tropoelastin, silicone, PNIPAAm-PEG, laminin, poly-L- ornithine, matrigel, and/or combinations thereof.

[0026] The above-mentioned coating increases the adhesion properties and provides other signals needed for growth and differentiation. For instance, the combined coating of poly-L-Ornithine with laminin on the tissue culture plastic or cell plate promotes the attachment of neural stem cells to it, and improves the efficiency of induced neural differentiation towards neurons, glial cells and other neural cells. Specifically, laminin is important for cell attachment as the cell receptors' integrins bind to multiple sites on laminin. As for poly-L-ornithine, this polymer activates the ERK signalling pathway which allows the differentiation of neural stem cells to occur.

[0027] In an embodiment, the cell plate/container, the cell plate lid and the cell culture partition may be preferably made of a plastic material, which can be thermoformed, or made by another suitable plastic forming process. The plastic materials, which are transparent to allow viewing of the media and any growing bacteria, are preferred. Materials that are also capable of retaining moisture are preferred. Examples of suitable materials include polyvinylchloride, polyethylene terephthalate, polytetrafluoroethylene, polyesters, polystyrenes, polycarbonates, and polyethylene, with polymers or without polymers. Selection of the appropriate plastic material is not crucial as long as the material is capable of retaining moisture and withstanding incubation conditions.

[0028] In an embodiment, when the partition or partitions are inserted, two or more compartments are created and each compartment is surrounded by limiting walls.

[0029] In an embodiment, the interior of the cell plate, which is in contact with the cell culture, and the partition may be coated with a membrane for efficiently block the transmission of light (from 400-700nm), allowing fluorescence detection in a simply and non-destructive manner.

[0030] In an embodiment, the bottom of the wells may be flat, round or conic.

[0031] In an embodiment, the bottom of the well may be flat when microscopic and optical measurements are wanted. Microscopic measurements include procedures such as immunofluorescence microscopy where certain cell components are labelled by antibodies, bright field microscopy in order to measure cell size and to determine cell geometry, and calcium imaging by fluorescence-based methods. Optical measurements include procedures such as cell viability tests performed by resazurin-based procedures and cytotoxicity assays by LDH-based procedures. [0032] In an embodiment, the bottom of the well may be round when agglutination assays are wanted. Agglutination assays include procedures such as diagnosis of pathogens in patient sera by a Widal test, blood typing and quantification of the relative concentration of viruses, bacteria or antibodies by hemagglutination assays.

[0033] In an embodiment the bottom of the well may be conic when precipitation, centrifugation or sample recovery is necessary. In adherent cell culture, these operations usually occur when cell pelleting or cell lysis are desired on wells. Cell pelleting and cell lysis are necessary steps for isolation of genomic DNA or quantification of protein by BCA assay.

[0034] In an embodiment, the cell plate now disclosed may be stackable, therefore, saving space when in storage or when in use, e.g. inside an incubator.

[0035] In an embodiment, the cell plate disclosed may be compatible with robotics, automated readers, and liquid handling systems.

[0036] In an embodiment, the cell plate disclosed may be used for drug screening and/or toxicity testing; in particular it may be used for a method of interacting more than one type of substances with a type of cell material, in a cell plate having a plurality of wells. This includes depositing a type of the cell material in the cell plate, agitating the cell plate for homogenising its spatial distribution, depositing a fluid medium, adding the partitions to the cell plate and adding the substance to the fluid medium of each tight-fluid well.

[0037] In an embodiment, the cell plate disclosed may be used for drug screening and/or toxicity of substances such as a chemical or a drug.

[0038] In an embodiment, the cell plate disclosed may be used for optimization of cell-based processes, such as cell growth, stem cell differentiation, virus/vaccine/VLP or protein production among others, by means of manipulation of concentrations of nutrients, growth factors, cytokines, small molecules, natural or synthetic substrates, by adding a type of cell material or by manipulating environmental conditions such as pH, oxygen concentration, temperature or other physical and chemical forces.

[0039] In an embodiment, the cell plate disclosed may be used for cell migration/invasion assays, in a cell plate having a plurality of wells. This includes adding the partitions to the cell plate to form the tight-fluid wells, depositing a type of the cell material in the wells created, depositing a fluid medium to each well, and remove one or more partitions to create cell-free gaps for migration/invasion assays.

[0040] In an embodiment, each partition is of a substantially rigid, fluid-impervious, typically thermoplastic material substantially chemically non-reactive with the fluids to be employed in the assays to be carried out using the cell plate now disclosed. The term "substantially rigid" as used herein means that the material resists deformation or warping under a light mechanical or thermal load, whose deformation would prevent maintenance of the substantially planar surface, although the material may be somewhat elastic.

[0041] In an embodiment, the cell plate depth, together with the area of the cell plate determines the volume capacity of each cell plate.

[0042] In an embodiment, the volume capacity of each well, once a multiwell plate is assembled, is determined by the well depth, together with the area of the respective well.

[0043] In an embodiment, each partition may comprise a sharp end/edge which may be straight, curved, serrated and/or combinations thereof and arranged in such a way that when the partition is introduced into the cell plate, fluid-tight wells are formed.

[0044] In an embodiment, each partition may comprise one, two or more blades arranged in such a way that when the partition is introduced into the cell plate, fluid-tight wells are formed.

[0045] In an embodiment, each partition may comprise blades with hollow grind, sabre grind, chisel grind, double bevel, convex grind forms and/or combinations thereof, arranged in such a way that when the partition is introduced into the cell plate, fluid-tight wells are formed.

[0046] In an embodiment, each partition may be a combination of two or more pieces, particularly the spine of the partition and the blades at its end, fused together or fixed together by mechanical joints, in such a way the partition is substantially rigid, fluid-impervious, and substantially chemically non-reactive with the fluids to be employed in the assays to be carried out using the cell plate now disclosed. The term "substantially rigid" as used herein means that the material resists deformation or warping under a light mechanical or thermal load, whose deformation would prevent maintenance of the substantially planar surface, although the material may be somewhat elastic.

[0047] In an embodiment, a partition may comprise one or more pins, wherein each pin is arranged to connect to a carved pin from a second partition.

[0048] The present disclosure also relates to a method for culturing cells comprising the following steps:

1) coating the plate, if necessary;

2) thawing a cryovial of cells or performing trypsinization of a cell culture, determining cell number and adding the needed cell number to the adequate volume of culture medium for the plate;

3) suspending cells in culture medium are seeded, in a single pipetting step, in an open space of the plate (partial if there are already partitions in the plate or total area of the plate if it is free of partitions);

4) letting cells adhere to the plate at certain environmental conditions, for example in an incubator; 5) after adhesion of cells, changing medium, whenever needed, is performed by aspirating a certain volume of exhaust medium and then discharging fresh medium to the plate;

6) sampling cells or media by inserting a specific partition or specific partitions, through the rails, for the area of sampling and for the number of replicates desired;

7) pressing the partitions against the plate to assure the area inside partitions are physically separated from the neighbouring areas;

8) sampling media and/or cells after previous physical separation of one or more divisions.

[0049] A rail according to the disclosure is an elongated feature that allows a partition to slide into insertion in the disclosed plate. For example, a rail may be an elongated channel for slidably receiving said partition or a pair of elongated protuberant ribs for slidably receiving said partition between the two ribs.

[0050] The present disclosure relates to a plate for multiwell cell culturing comprising: a bottom surface;

a plurality of sidewalls;

at least one rail for the insertion of a first fluid-tight partition;

wherein the at least one rail is arranged on a sidewall; and

the insertion of the first fluid-tight partition into the at least one rail is configured to generate a plurality of fluid-tight wells.

[0051] In an embodiment, the plate for multiwell cell culturing now disclosed comprises: a bottom surface; a plurality of sidewalls; at least a first rail and a second rail, for the insertion of a first fluid-tight partition; wherein the first rail is arranged on a sidewall opposite to the sidewall of the second rail; the first rail and the second rail face each other; and the insertion of the first fluid-tight partition into the first rail and the second rail is configured to generate a plurality of fluid-tight wells.

[0052] In an embodiment, the at least one rail may be a channel, preferably the rails may be channels.

[0053] In an embodiment, the at least one rail may have two ribs, preferably each rail may have two ribs.

[0054] In an embodiment, the at least one rail may be run the full height of the respective sidewall or the at least one rail may be run the partial height of the respective sidewall.

[0055] In an embodiment, at least one rails may be run the full height of the respective sidewall or at least one rails may be run the partial height of the respective sidewall. [0056] In an embodiment, the plate may comprise at least a first rail and a second rail, for the insertion of the first fluid-tight partition, wherein the first rail is in a sidewall opposite to the sidewall of the second rail and wherein said rails face each other.

[0057] In an embodiment, the plate now disclosed may comprise at least a fluid-tight partition, preferably a first fluid-tight partition, wherein the fluid-tight partition has a sharp edge configured to contact with the bottom surface of the plate.

[0058] In an embodiment, the first fluid-tight partition may comprise one or more pins configured to couple to a second fluid-tight partition or to further fluid-tight partitions.

[0059] In an embodiment, the first fluid-tight partition and the second fluid-tight partition may have a sharp edge configured to contact with the bottom surface of the plate.

[0060] In an embodiment, any of the fluid-tight partitions may have a top edge below the height of the plate.

[0061] In an embodiment, the at least two fluid-tight wells may share a common bottom surface.

[0062] In an embodiment, the plurality of fluid-tight wells may be 2, 4, 6, 12, 24, 96 or 384 fluid-tight wells.

[0063] In an embodiment, the sidewalls may be pairwise, each sidewall pair is placed in front of each other, and a rail is arranged on each of said sidewalls, such that a plurality of fluid-tight wells are obtained when a fluid-tight partition is inserted into said rails, the fluid-tight partition extending between the two sidewalls of each sidewall pair.

[0064] In an embodiment, the plate may comprise four sidewalls.

[0065] In an embodiment the bottom surface of the plate bottom may be flat.

[0066] In an embodiment, the bottom surface of the plate bottom may be patterned of round depressions or patterned of conic depressions.

[0067] In an embodiment, the plate may be made of polyvinylchloride, polyethylene terephthalate, polytetrafluoroethylene, polyesters, polystyrenes, polycarbonates, and polyethylene.

[0068] In an embodiment, the any of the fluid-tight partitions may be made of polyvinylchloride, polyethylene terephthalate, polytetrafluoroethylene, polyesters, polystyrenes, polycarbonates, and polyethylene.

[0069] In an embodiment, the plate may comprise a lid for closing the plate.

[0070] In an embodiment, the lid may be made of polyvinylchloride, polyethylene terephthalate, polytetrafluoroethylene, polyesters, polystyrenes, polycarbonates, and polyethylene. [0071] In an embodiment, the bottom surface of the plate may be coated with collagen, poly-L-Lysine, poly-D-Lysine, fibronectin, vitronectin, hyaluronan, fibroin silk, tropoelastin, silicone, PNIPAAm-PEG, laminin, poly-L-ornithine, matrigel, and/or combinations thereof.

[0072] In an embodiment, any of the fluid-tight partition may be coated with collagen, poly-L-Lysine, poly-D-Lysine, fibronectin, vitronectin, hyaluronan, fibroin silk, tropoelastin, silicone, PNIPAAm-PEG, laminin, poly-L-ornithine, matrigel, and/or combinations thereof.

[0073] The present disclosure also relates to the use of the plate now disclosed for multiwell cell culturing.

Brief description of the drawings

[0074] The following figures provide preferred embodiments for illustrating the description and should not be seen as limiting the scope of the disclosure.

[0075] Figure 1: Schematic representation of a top view of a cell plate (1) with rails (3).

[0076] Figure 2: Schematic representation of a top view of a cell plate (1) with rails (3).

[0077] Figure 3: Schematic representation of a top view of a cell plate (1) with rails and partitions (4).

[0078] Figure 4: Schematic representation of a top view of a cell plate (1) with rails and partitions (4).

[0079] Figure 5: Schematic representation of a lateral view of an open cell plate (1).

[0080] Figure 6: Schematic representation of a lateral view of a closed cell plate (1).

[0081] Figure 7: Schematic representation of a lateral view of an open cell plate (1) with rails (3).

[0082] Figure 8: Schematic representation of a lateral view of an open cell plate (1) comprising cells (7), and wherein a partition (4) is being inserted.

[0083] Figure 9: Schematic representation of a lateral view of the cell plate (1) wherein a plurality of cells (7) are deposited in the bottom (6) of the cell plate (1), covered by a cell medium and wherein each partition (4) is placed between a pair of rails (3).

[0084] Figure 10 represents a lateral male partition of level 1 (LM 1).

[0085] Figure 11 represents a lateral partition with a male pin of level 2 and a female pin receptor of level 1 (LM2F1).

[0086] Figure 12 represents an in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a male pin of level 1 (IM3F2-M1).

[0087] Figure 13 represents an in-between partition in which on one side it has a male pin of level 2 and a female pin receptor of level 1, and on the other side a female pin receptor of level 3 (IM2F1-F3). [0088] Figure 14 represents a lateral female partition of level 3 (LF3).

[0089] Figure 15 represents a lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2).

[0090] Figure 16 represents a lateral to lateral partition with two male pins of level 2 (LLM2x2).

[0091] Figure 17 represents an in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a female pin receptor of level 3 (IM3F2-F3).

[0092] Figure 18 represents a possible formation of 4 wells, in a well-by-well manner, in a cell plate capable of accommodating 12 wells, by insertion of specific partitions into the cell plate.

[0093] Figure 19 represents a possible formation of 6 wells, in a 3 replicates per time-point manner, in a cell plate capable of accommodating 12 wells, by insertion of specific partitions into the cell plate.

Detailed description

[0094] Figure 1 represents a top view of a cell plate (1) of the present disclosure displaying four lateral walls (2), wherein the lateral walls are pairwise, and each lateral wall pair wised is placed in front of each other. Two of the four lateral walls, placed in front of each other, comprise a plurality of rails (3) for the insertion of partitions.

[0095] Figure 2 represents a top view of the cell plate (1) now disclosed wherein the four lateral walls (2) comprise a plurality of rails (3) for the insertion of partitions.

[0096] Figure 3 and Figure 4 represent a top view of the cell plate (1) now disclosed wherein several partitions (4) are inserted, therefore creating several independent and sealed compartments.

[0097] Figure 5 and Figure 6 represent a lateral view of the cell plate (1) now disclosed comprising a lid (5) and a bottom (6), wherein in Figure 5 the cell plate (1) is open and in Figure 6 the cell plate (1) is closed.

[0098] Figure 7 represents a lateral view of the open cell plate (1) now disclosed comprising a plurality of rails (3).

[0099] Figure 8 represents a lateral view of the cell plate (1) now disclosed wherein a plurality of cells (7) are deposited in the bottom (6) of the cell plate (1) and covered by a cell medium. Figure 8 also represents a partition (4) wherein said partition (4) may comprise a sharp end (8).

[00100] Figure 9 represents a lateral view of the cell plate (1) wherein a plurality of cells (7) are deposited in the bottom (6) of the cell plate (1), covered by a cell medium and wherein each partition (4) is placed between a pair of rails (3).

[00101] Examples on possible partitions that may be used in the cell plate now disclosed. [0Q102] Figure 10A represents a front side of a lateral male partition of level 1 (LMl), wherein (9) represents the height of the piece, that is the lateral male partition of level 1 (LMl); (10) is the internal side of the well; (11) is the width of the partition to be coupled to this LMl partition; (12) is the height of level 1; Figure 10B represents a first side of the lateral male partition of level 1 (for example the right side); and Figure IOC represents a second side of the lateral male partition of level 1 (for example the left side).

[00103] Figure 11A represents a lateral partition with a male pin of level 2 and a female pin receptor of level 1 (LM2F1), wherein (9) represents the height of the piece, that is the lateral partition with a male pin of level 2 and a female pin receptor of level 1 (LM2F1); (10) is the internal side of the well; (11) is the width of the partitions to be coupled to this LM2F1 partition; (12) represents the height of level 1; (13) represents the height of level 2; Figure 11B represents a first side of the lateral partition with a male pin of level 2 and a female pin receptor of level 1 (LM2F1) (for example the right side); and Figure 11C represents a second side of the lateral partition with a male pin of level 2 and a female pin receptor of level 1 (LM2F1) (for example the left side).

[00104] Figure 12A represents an in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a male pin of level 1 (IM3F2-M1), wherein (10) is the internal side of the well; (11) is the width of the partitions to be coupled to this IM3F2-M 1 partition; (12) is the height of level 1; (13) is the height of level 2; (14) is the height of level 3; Figure 12B represents a first side of the in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a male pin of level 1 (IM3F2-M1) (for example the right side); and Figure 12C represents a second side of the in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a male pin of level 1 (IM3F2-M1) (for example the left side).

[00105] Figure 13A represents an in-between partition in which on one side it has a male pin of level 2 and a female pin receptor of level 1, and on the other side a female pin receptor of level 3 (IM2F1-F3), wherein (10) is the internal side of the well, (11) is the width of the partitions to be coupled to this IM2F1- F3 partition; (12) is the height of level 1; (13) is the height of level 2; (14) is the height of level 3; Figure 13B represents a first side of the in-between partition in which on one side it has a male pin of level 2 and a female pin receptor of level 1 (for example the right side), and on the other side a female pin receptor of level 3 (IM2F1-F3); and Figure 13C represents a second side of the partition in which on one side it has a male pin of level 2 and a female pin receptor of level 1, and on the other side a female pin receptor of level 3 (IM2F1-F3).

[00106] Figure 14A represents a lateral female partition of level 3 (LF3), wherein (9) represents the height of the piece, that is the lateral female partition of level 3 (LF3); (10) represents the internal side of the well; (11) represents the width of the partition to be coupled to this LF3 partition; (14) represents the height of level 3; Figure 14B represents a first side of the lateral female partition of level 3 (LF3) (for example the right side); and Figure 14C represents a second side of the lateral female partition of level 3 (LF3) (for example the left side).

[00107] Figure 15A represents a lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2), wherein (9) represents the height of the piece, that is the lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2); (10) represents the internal side of the well; (11) represents the width of the partitions to be coupled to this LM3F2 partition; (13) is the height of level 2; (14) represents the height of level 3; Figure 15B represents a first side of the lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2) (for example the right side); and Figure 15C represents a second side of the a lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2) (for example the left side).

[00108] Figure 16A represents a lateral to lateral partition with two male pins of level 2 (LLM2x2) wherein

(9) represents the height of the piece, that is the lateral partition with two male pins of level 2 (LLM2x2);

(10) represents the internal side of the well; (11) represents the width of the partitions to be coupled to this LLM2x2 partition; (13) represents the height of level 2; Figure 16B represents a first side of the lateral to lateral partition with two male pins of level 2 (LLM2x2) (for example the right side) and Figure 16C represents a second side of the lateral to lateral partition with two male pins of level 2 (LLM2x2) (for example the left side).

[00109] Figure 17A represents an in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a female pin receptor of level 3 (IM3F2-F3) wherein (9) represents the height of the piece, that is the in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a female pin receptor of level 3 (IM3F2-F3); (10) represents the internal side of the well; (11) represents the width of the partitions to be coupled to this IM3F2-F3 partition; (13) represents the height of level 2 and (14) represents the height of level 3; Figure 17B represents a first side of the in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a female pin receptor of level 3 (IM3F2-F3) (for example the right side) and Figure 17C represents a second side of the in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a female pin receptor of level 3 (IM3F2-F3) (for example the left side).

[00110] Examples on how to assembly the partitions in the cell plate now disclosed.

[00111] In an embodiment, specific partitions may be combined in the cell plate now disclosed, as indicated in Figure 18. Figure 18, which represents a formation of 4 wells, in a well-by-well manner, in a cell plate capable of accommodating up to 12 wells. In this embodiment, partitions of the following type are sequentially combined: a lateral male partition of level 1 (LM 1), a lateral partition with a male pin of level 2 and a female pin receptor of level 1 (LM2F1), an in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a male pin of level 1 (IM3F2- M l), a lateral partition with a male pin of level 2 and a female pin receptor of level 1 (LM2F1), a lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2), lateral male partition of level 1 (LM 1), an in-between partition in which on one side it has a male pin of level 2 and a female pin receptor of level 1, and on the other side a female pin receptor of level 3 (IM2F1-F3).

[00112] In an embodiment, specific partitions may also be combined in the cell plate now disclosed, as indicated in Figure 19. Figure 19 represents a formation of 6 wells, in a 3 replicates per time-point manner, in a cell plate capable of accommodating up to 12 wells, by insertion of specific partitions into the cell plate. In this embodiment, partitions of the following type are sequentially combined: a lateral to lateral partition with two male pins of level 2 (LLM2x2), a lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2), a lateral partition with a male pin of level 3 and a female pin receptor of level 2 (LM3F2), a lateral to lateral partition with two male pins of level 2 (LLM2x2), an in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a female pin receptor of level 3 (IM3F2-F3), an in-between partition in which on one side it has a male pin of level 3 and a female pin receptor of level 2, and on the other side a female pin receptor of level 3 (IM3F2-F3), a lateral to lateral partition with two male pins of level 2 (LLM2x2).

[00113] In an embodiment, one or more partitions (4) may be added to the cell plate in order to create two or more compartments, respectively.

[00114] In an embodiment, each cell plate may be a four well plate, for example, with about 3.15 cm x 3.15 cm and 1.1 cm deep, and the wells being preferably arranged in a 2 by 2 regular rectangular array.

[00115] In an embodiment, each cell plate may be a six well plate, for example, with about 4.12 cm x 4.05 cm and 1.77 cm deep, and the wells being preferably arranged in a 2 by 3 regular rectangular array.

[00116] In an embodiment, each cell plate may be an eight well plate, for example, with about 4.12 cm x 2.96 cm and 1.77 cm deep, and the wells being preferably arranged in a 2 by 4 regular rectangular array.

[00117] In an embodiment, each cell plate may be a twelve well plate, for example, with about 2.64 cm x 2.96 cm and 1.77 cm deep, and the wells being preferably arranged in a 4 by 3 regular rectangular array.

[00118] In an embodiment, each cell plate may be a twenty-four well plate, for example, with about 1.91 cm x 1.87 cm and 1.77 cm deep, and the wells being preferably arranged in a 6 by 4 regular rectangular array.

[00119] In an embodiment, each cell plate may be a ninety-six well plate, for example, with about 0.80 cm x 0.78 cm and 1.1 cm deep, and the wells being preferably arranged in a 12 by 8 regular rectangular array. [00120] In an embodiment, each cell plate may accommodate a pre-arranged partitions array in order to form, in a single-step, a multi-well plate. The pre-arranged partitions array may be divided in any multi- well format such as 6, 12, 24, 96 or 384 wells and placed at once in the rails.

[00121] In an embodiment, this disclosure may be applied to any multi-well format with any number of wells per cell plate.

Example 1

[00122]Cells and culture medium can be seeded on a culture plate in order to perform several metabolic flux analysis, for example, in order to perform a ¹³ C metabolic flux analysis. In this metabolic flux analysis study, the ¹³ C metabolic flux analysis study, two types of data are usually needed: 1) secretion and uptake of nutrients from the culture medium over cell culture time; and 2) profile of labelling of intracellular metabolites over culture time after an addition of ¹³ C-labelled nutrient to the culture medium. These data require sampling of culture media and of cells, respectively.

[00123]The number of time-points and the number of biological replicates per time-point should be chosen so that the culture plate may be dividable in the desired total number of wells. The total number of wells is determined by multiplying time-points with replicates. For example, if eight time-points and three biological replicates per time-point are desired, the plate should have the necessary rails and the number of partitions attached for creating a 24 well plate, all with the same area.

[00124] For the first time-point, a partition may be inserted across the width of the culture plate. Then, two other small inserts may be inserted in perpendicular crossing the border of the culture plate to the partition just inserted, in order to form three biological replicates. These three wells would be thus physically separated and the area of each one would be equal to area of the culture plate without partition dividing by 24 final wells. Medium and cells of each well can then be sampled well by well.

[00125]This procedure may be repeated for the next seven time-points, each with three biological replicates.

[00126] Each biological replicate may have shared the same culture medium and cells seeded in the beginning of the culture. Moreover, each sample of a given time-point, may have shared the same secretion or uptake of nutrients and the same intracellular labelling of metabolites as of the previous time-points.

[00127] In this manner, concentrations of nutrients in the media and profiles of labelling of intracellular metabolites come associated with a rather small biological variation. Moreover, this is then reflected in the estimation of extracellular and intracellular fluxes, which will present small relative deviations of calculated flux values.

Example 2 [00128]Cells and culture medium can be seeded on a culture plate in order to perform drug screening or toxicity testing. Cells attach to the culture plate and grow until reaching a confluent layer of cells. Then, a pre-arranged set of partitions may be inserted in the cell confluent culture plate in order to form a 96 well plate. After assuring that the partitions have created 96 physically separated wells, the administration of drugs for drug screening or toxicity testing may begin. In this manner, the comparison of efficacy or toxicity of drugs may become facilitated as all wells originated from the same confluent cell monolayer and from the same medium.

Example 3

[00129]Culture medium and 3D aggregates can be seeded on a culture plate in order to perform drug screening or toxicity testing. The cell plate is agitated to induce a homogeneous spatial distribution of 3D aggregates in the liquid medium. Then, a pre-arranged set of partitions may be inserted in the culture plate with 3D aggregates in suspension in order to form a 96 well plate. After assuring that the partitions have created 96 physically separated wells, the administration of drugs for drug screening or toxicity testing may begin. In this manner, the comparison of efficacy or toxicity of drugs may become facilitated as all wells contain the same medium and approximately share the same probability distribution of 3D aggregates as of the whole population in the cell plate.

Example 4

[00130]Cells and culture medium can be seeded on a culture plate in order to perform optimization of conditions for co-culture of cells. Cells, for example, mesenchymal stem cells (MSC), attach to the culture plate and grow until reaching a confluent layer of cells. Then, cells in suspension may be added to the culture plate, in particular, hematopoietic stem cells (HSC). To the culture plate with a co-culture, a prearranged set of partitions may be inserted in order to form 24 wells. After assuring the partitions created 24 physically separated, different experimental conditions may be applied to different wells in order to optimize one or more cell parameters. In this manner, the power of design of experiments for multivariate optimizations may be better harnessed using a culture plate in which each well share the same cells and medium.

Example 5

[00131]Cells and culture medium can be seeded on a culture plate in order to perform migration or cell invasion assays. A pre-arranged set of partitions may be inserted in the culture plate in order to form a 24 well plate. Cells, for example, mesenchymal stem cells (MSC), are seeded into each well, attach to the culture plate and grow until reaching a confluent layer of cells in each well. Some partitions are then removed in order to form a 12 well plate, leaving gaps of cell-free culture plate and cell migration may be then evaluated by microscopy. [00132]Throughout the description and claims the word "comprise" and variations of the word, are not intended to exclude other technical features, additives, components, or steps. Additional objectives, advantages and features of the solution will become apparent to those skilled in the art upon examination of the description or may be learned by practice of the solution.

[00133]The term "comprising" whenever used in this document is intended to indicate the presence of stated features, integers, steps, components, but not to preclude the presence or addition of one or more other features, integers, steps, components or groups thereof.

[00134] It will be appreciated by those of ordinary skill in the art that unless otherwise indicated herein, the particular sequence of steps described is illustrative only and can be varied without departing from the disclosure. Thus, unless otherwise stated the steps described are so unordered meaning that, when possible, the steps can be performed in any convenient or desirable order.

[00135]The disclosure should not be seen in any way restricted to the embodiments described and a person with ordinary skill in the art will foresee many possibilities to modifications thereof.

[00136]The above-described embodiments are combinable.