WO2021207817A1 - Cell culture inserts containing hydrogel and method of use - Google Patents
Cell culture inserts containing hydrogel and method of use Download PDFInfo
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- WO2021207817A1 WO2021207817A1 PCT/BR2021/050159 BR2021050159W WO2021207817A1 WO 2021207817 A1 WO2021207817 A1 WO 2021207817A1 BR 2021050159 W BR2021050159 W BR 2021050159W WO 2021207817 A1 WO2021207817 A1 WO 2021207817A1
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- cell culture
- hydrogel
- fact
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- insert containing
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- 238000004113 cell culture Methods 0.000 title claims abstract description 62
- 239000000017 hydrogel Substances 0.000 title claims abstract description 52
- 238000000034 method Methods 0.000 title claims abstract description 11
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/02—Membranes; Filters
- C12M25/04—Membranes; Filters in combination with well or multiwell plates, i.e. culture inserts
Definitions
- the present invention relates with cell culture inserts containing a layer of hydrogel for use in cell culture on air-gel interfaces in order to promote the formation of constructs similar to epithelial tissues.
- the present invention also deals with the method of using such cell culture inserts.
- the patent US4871674 discloses an insert containing discontinuous projections to hang it in an upper circumferential portion and one membrane filter in a lower portion, being characterized by having triangular conical ribs provided continuously underneath the discontinuous projection parts.
- the US5026649 patent discloses an apparatus forgrowing tissues "in vitro", which allows a gradient of nutrient concentration to develop through a permeable membrane to which a tissue sample is attached.
- the permeable membrane is attached to the lower end of a tubular support which, in turn, is suspended by a flange connected to its upper end at the top of a vat containing nutrients.
- the vat is usually part of a tissue culture plate set.
- the flange positions the support and the membrane centrally in the bowl, in order to avoid capillary action in the space between the bowl and the support.
- the configuration of the support and its cooperation with the cover of the plate assembly also prevent the support and the membrane from floating in the nutrient solution of the vat.
- the openings in the holder provide access to a pipette to add and remove fluid from the space between the bowl and the membrane holder and from the space below the membrane.
- US5578492 discloses an insert containing a membrane for cell culture and a device for separating the membrane to facilitate the use of conventional instrumentation for further examination.
- the cell culture insert is provided with a microporous membrane forming the bottom wall as the cell culture surface of the insert.
- the insert includes a support mechanism to hold it suspended in a certain position on a plate with several culture vessels and a mechanism to separate the support from the membrane supporting portion of the insert, once the desired degree of cell culture is achieved has been reached.
- US5652142 discloses an apparatus for culturing cells or tissues "in vitro" that allows the development of a gradient of nutrient concentration through a permeable membrane to which a tissue sample is attached.
- the permeable membrane is attached to the lower end of a cell culture insert which, in turn, is supported by a flange connected at its upper end at the top of a vat containing nutrients.
- the restricted movement of the cell culture insert in the vat can be facilitated by projections extending outside the surface of the outer wall of the insert, in order to avoid capillary action in the space between the vat and the insert.
- patent US7598076 discloses a cell culture insert comprising a cup-shaped wall, a base consisting of a filtration membrane, projecting support arms that are distributed around the periphery of the top, and lateral spacers for vertical and horizontal orientation in a vat containing liquid culture medium on a cell culture plate.
- the spacers are distributed around the periphery of the cell culture insert and have different radial lengths in such a way that a large feed opening and several smaller openings are formed.
- the aforementioned documents generally describe a plastic cell culture insert comprising a side wall bounded by an upper-end opening and lower-end opening, with a flat permeable membrane attached to the lower-end opening.
- these cell inserts are used to allow the parallel culture of two types of cells, one on the cell culture plate and the other on the cell culture insert, so that the interaction between these two types of cells can be studied without the real physical contact between them.
- the present invention deals generally with cell culture inserts containing a hydrogel layer for use in cell culture on air-gel interfaces to promote the formation of epithelial tissue-like constructions and methods of using these inserts.
- Epithelial tissues grown on air-gel interfaces according to the present invention represent a microenvironment more similar to “in vivo” when compared to monolayers in conventional two- dimensional culture systems or air-liquid interfaces.
- Epithelial cells are present in different tissues of the body, such as the skin, the digestive tract, the pulmonary tract, the urinary tract, and the glands. In all of these tissues, epithelial cells rest on a layer of extracellular matrix that acts as a support on which the epithelium can proliferate.
- the hydrogel layer in the cell culture insert according to the present invention efficiently mimics the microenvironment in which the cells are naturally arranged.
- the present invention therefore, deals with three-dimensional cell culture inserts containing a hydrogel layer for cell culture on air-gel interfaces to promote the formation of epithelial tissue-like constructions and methods of using these inserts.
- Said inserts comprise a hollow chamber with two distinct regions - upper and lower, a hydrogel filling, and a support capsule.
- Figures 1 A-1 E - illustrate a reconstructed schematic representation in 3D of the cell culture insert object of the present invention without the hydrogel.
- Figure 2 - illustrates a schematic representation of the cell culture insert object of the present invention showing the air-gel interfaces.
- Figure 3 - illustrates a schematic representation of the methodological sequence in which the insert is configured to promote the formation of constructs similar to epithelial tissues.
- Figures 1A-1E show in three-dimensional schematic representation the main characteristics of the cell culture insert according to the present invention without the hydrogel.
- the design of the insert holder can also be visualized.
- Figure 1A a top view of the insert
- Figure 1B a bottom view of the insert
- Figure 1C a side view of the insert
- Figure 1D a cross-sectional view of the insert
- Figure 1E a general perspective view of the insert.
- the cell culture insert according to the present invention comprises a hollow chamber with two distinct portions, an upper portion (1) and a lower portion (2), and a support base (3).
- the hollow chamber has the upper portion (1) with an open end at the top and the lower portion (2) with a perforated and closed-end at the bottom (4).
- the upper portion (1) of the said hollow chamber has sloping side walls (11), as the diameter of the upper portion (12) is greater than the diameter of the lower portion (13).
- one or more flaps (14) directed towards the interior of the hollow chamber are provided, in order to facilitate the manipulation of the cell culture insert.
- the lower portion (2) of the chamber contains side walls (21), preferably perforated, in case of which provided with multiple holes that connect the external and internal surfaces of the same.
- each hole has a diameter ranging from 0.1 pm to 10,000 pm, preferably from 1 pm to 7,500 pm, more preferably from 10 pm to 5,500 pm and even more preferably from 100 pm to 2,500 pm.
- a hydrogel plug (6) fills the entire volume of the lower portion (2) of the hollow chamber and on which the cells (7) are arranged.
- the bottom (4) of the hollow chamber is drilled through multiple holes (5) that connect the external and internal surfaces.
- each hole has a diameter ranging from 0.1 pm to 10,000 pm, preferably from 1 pm to 7,500 pm, more preferably from 10 pm to 5,500 pm and even more preferably from 100 pm to 2,500 pm.
- the hollow chamber (1 , 2) has a support base (3) provided with at least two inclined legs (31 , 32) with spacing between them in order to allow the flow of fluids from the culture medium (8) towards the bottom (4) with perforations (5) of the hollow chamber (1 , 2), thus reaching the hydrogel layer (6) on which the cells (7) rest.
- the tip of each leg (31 , 32) touches the bottom edge of the said vessel (9) in order to stabilize the cell culture insert.
- the outer dimensions of the cell culture insert object of the present invention are sufficiently smaller than the internal dimensions of the tissue culture vessel to allow the insert to be completely inserted into the tissue culture vessel.
- the cell culture insert according to the present invention can be sized to be adapted for use with different tissue culture vessels and vats of different sizes and geometric configurations.
- the hydrogel used in the cell culture insert according to the present invention can be any of the synthetic type, such as, for example, those produced in polyethylene glycol, polyacrylic acid, polyvinyl alcohol, and polypeptides, but not limited to them.
- a natural hydrogel can also be used, such as, for example, fibrin, collagen, gelatin, hydrogels derived from the decellularized extracellular matrix, hyaluronic acid, alginate, agarose, nanocellulose and modified natural polymers, but not limited to them. Mixtures of these hydrogels or compositions thereof can also be used.
- C carbonyls
- CO2H carboxylic acids
- CO2R esters
- NH2 amines
- the hydrogel for use in the cell culture insert according to the present invention can also have surface topography characteristics, including columns, pits, grooves, hemispheres, and grids, among other possible ones.
- the present invention also deals with a method of using the cell culture insert according to the present invention, comprising introducing target cells on top of the hydrogel (6) and a cell culture medium (8) in the vessel (9) where the insert is incorporated to grow the target cells in an air-gel interface.
- the method of use according to the present invention includes chemical, biochemical, or biological compounds that are added to the medium contained in the vessel (9), so that they move from said vessel (9) to the hydrogel (6), pass through the hydrogel (6) and reach cell culture (7).
- Biological agents for example, prions, viruses, bacteria, and protozoa, without limitation, are added to the medium (8) contained in the vessel (9), so that they move from the vessel (9) to the hydrogel (6), passing through the hydrogel (6) and reaching the cell culture (7).
- said biological agents can be added directly to the cells (7) that grow on top of the hydrogel (6), the effects of such treatments being evaluated on the target cells (7).
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
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Abstract
The present invention deals with cell culture inserts containing a hydrogel layer for use in cell culture on air-gel interfaces in order to promote the formation of constructions similar to epithelial tissues, as well as deals with the method of using these cell culture inserts. The insert according to the 5 present invention comprises a hollow chamber with two distinct portions, an upper portion (1) and a lower portion (2), and a support base (3), said upper portion (1) having an open end at the top and said lower portion (2) having side walls (21) and being closed at the bottom (4) perforated (5), provided with a hydrogel plug (6) on which the cells (7) are arranged.
Description
CELL CULTURE INSERTS CONTAINING HYDROGEL AND METHOD OF USE
FIELD OF THE INVENTION
The present invention relates with cell culture inserts containing a layer of hydrogel for use in cell culture on air-gel interfaces in order to promote the formation of constructs similar to epithelial tissues. The present invention also deals with the method of using such cell culture inserts.
BACKGROUND OF THE INVENTION
Pre-clinical studies, like animal models, have serious problems, such as its high biological, ethical and financial cost. Thus, most of the research conducted in the laboratory is based on the use of “in vitro” models that seek to mimic and replicate structures observed in living organisms, notably for the development of new drugs. In general, cell cultures are performed with the aid of devices made of plastic material, or glass, in two-dimensional models, which are not yet suitable for reproducing the natural microenvironment in which cells are originally found in living organisms. Therefore, the use of tissue engineering has been an alternative to traditional two-dimensional cell culture and “in vivo” models when using biomimetic materials and cell lines to mimic tissues in a three-dimensional environment. The use of three-dimensional cell culture models demonstrates advantages for the study of cells, notably to mimic and study different pathologies
Conventional cell culture inserts and devices are widely described by the state of the art, for example, US Patents US4871674 by Matsui, Hirai and Nakamura, 1989; US5026649 by Lyman, Mathus and Root, 1991 ; US5578492 by Fedun, 1996); US5652142 by Barker et al., 1997; and US7598076B2 by Wedell and Matthes, 2009.
The patent US4871674 discloses an insert containing discontinuous projections to hang it in an upper circumferential portion and one membrane filter in a lower portion, being characterized by having triangular conical ribs provided continuously underneath the discontinuous projection parts.
The US5026649 patent discloses an apparatus forgrowing tissues "in vitro", which allows a gradient of nutrient concentration to develop through a permeable membrane to which a tissue sample is attached. The permeable membrane is attached to the lower end of a tubular support which, in turn, is suspended by a flange connected to its upper end at the top of a vat containing nutrients. The vat is usually part of a tissue culture plate set. The flange positions the support and the membrane centrally in the bowl, in order to avoid capillary action in the space between the bowl and the support. The configuration of the support and its cooperation with the cover of the plate assembly also prevent the support and the membrane from floating in the nutrient solution of the vat. The openings in the holder provide access to a pipette to add and remove fluid from the space between the bowl and the membrane holder and from the space below the membrane.
US5578492 discloses an insert containing a membrane for cell culture and a device for separating
the membrane to facilitate the use of conventional instrumentation for further examination. The cell culture insert is provided with a microporous membrane forming the bottom wall as the cell culture surface of the insert. The insert includes a support mechanism to hold it suspended in a certain position on a plate with several culture vessels and a mechanism to separate the support from the membrane supporting portion of the insert, once the desired degree of cell culture is achieved has been reached.
US5652142 discloses an apparatus for culturing cells or tissues "in vitro" that allows the development of a gradient of nutrient concentration through a permeable membrane to which a tissue sample is attached. The permeable membrane is attached to the lower end of a cell culture insert which, in turn, is supported by a flange connected at its upper end at the top of a vat containing nutrients. The restricted movement of the cell culture insert in the vat can be facilitated by projections extending outside the surface of the outer wall of the insert, in order to avoid capillary action in the space between the vat and the insert.
Finally, patent US7598076 discloses a cell culture insert comprising a cup-shaped wall, a base consisting of a filtration membrane, projecting support arms that are distributed around the periphery of the top, and lateral spacers for vertical and horizontal orientation in a vat containing liquid culture medium on a cell culture plate. The spacers are distributed around the periphery of the cell culture insert and have different radial lengths in such a way that a large feed opening and several smaller openings are formed.
The aforementioned documents generally describe a plastic cell culture insert comprising a side wall bounded by an upper-end opening and lower-end opening, with a flat permeable membrane attached to the lower-end opening. In most cases, these cell inserts are used to allow the parallel culture of two types of cells, one on the cell culture plate and the other on the cell culture insert, so that the interaction between these two types of cells can be studied without the real physical contact between them.
Another possible use of these cell culture inserts is to grow cells at air-liquid interfaces. In such experiments, cells are grown on top of the flat permeable membrane attached to the lower end of the insert, so that they come into contact with the cell culture medium on its lower surface and with air on its upper surface. This configuration is particularly interesting for the culture of epithelial cells, which require being in contact with the air to mature and express their characteristic phenotype. The permeable membrane present in conventional cell culture inserts, however, exposes the cells to a microenvironment that is very different from that found in the human body. This leads to a low success rate among compounds subjected to preclinical and clinical tests, wasting time and resources that could be applied to the development of compounds with a greater chance of success. Therefore, there is still a need to provide new and improved cell culture inserts capable of offering
cells and tissues a mimetic microenvironment to that found in the human body and, thus, increase the potential for translation of the results found in basic research.
BRIEF DESCRIPTION OF THE INVENTION The present invention deals generally with cell culture inserts containing a hydrogel layer for use in cell culture on air-gel interfaces to promote the formation of epithelial tissue-like constructions and methods of using these inserts.
Epithelial tissues grown on air-gel interfaces according to the present invention represent a microenvironment more similar to “in vivo” when compared to monolayers in conventional two- dimensional culture systems or air-liquid interfaces.
Epithelial cells are present in different tissues of the body, such as the skin, the digestive tract, the pulmonary tract, the urinary tract, and the glands. In all of these tissues, epithelial cells rest on a layer of extracellular matrix that acts as a support on which the epithelium can proliferate. The hydrogel layer in the cell culture insert according to the present invention efficiently mimics the microenvironment in which the cells are naturally arranged.
The present invention, therefore, deals with three-dimensional cell culture inserts containing a hydrogel layer for cell culture on air-gel interfaces to promote the formation of epithelial tissue-like constructions and methods of using these inserts. Said inserts comprise a hollow chamber with two distinct regions - upper and lower, a hydrogel filling, and a support capsule.
BRIEF DESCRIPTION OF THE DRAWINGS The inserts according to the present invention can be better understood with the description of the attached figures, which in a way only illustrative and not limiting its scope, represent:
Figures 1 A-1 E - illustrate a reconstructed schematic representation in 3D of the cell culture insert object of the present invention without the hydrogel.
Figure 2 - illustrates a schematic representation of the cell culture insert object of the present invention showing the air-gel interfaces.
Figure 3 - illustrates a schematic representation of the methodological sequence in which the insert is configured to promote the formation of constructs similar to epithelial tissues.
DETAILED DESCRIPTION OF THE INVENTION Figures 1A-1E show in three-dimensional schematic representation the main characteristics of the cell culture insert according to the present invention without the hydrogel. In addition to the general design of the insert, the design of the insert holder can also be visualized. Thus, we have in Figure 1A a top view of the insert, in Figure 1B a bottom view of the insert, in Figure 1C a side view of the insert, in Figure 1D a cross-sectional view of the insert, and in Figure 1E a general perspective view of the insert.
More specifically referring to Figures 2 and 3, it is observed that the cell culture insert according to the present invention comprises a hollow chamber with two distinct portions, an upper portion (1) and a lower portion (2), and a support base (3).
The hollow chamber has the upper portion (1) with an open end at the top and the lower portion (2) with a perforated and closed-end at the bottom (4). The upper portion (1) of the said hollow chamber has sloping side walls (11), as the diameter of the upper portion (12) is greater than the diameter of the lower portion (13). At the top of the said upper portion (1), at the open end, one or more flaps (14) directed towards the interior of the hollow chamber are provided, in order to facilitate the manipulation of the cell culture insert.
The lower portion (2) of the chamber contains side walls (21), preferably perforated, in case of which provided with multiple holes that connect the external and internal surfaces of the same. In general, as an example, it is preferred that each hole has a diameter ranging from 0.1 pm to 10,000 pm, preferably from 1 pm to 7,500 pm, more preferably from 10 pm to 5,500 pm and even more preferably from 100 pm to 2,500 pm. A hydrogel plug (6) fills the entire volume of the lower portion (2) of the hollow chamber and on which the cells (7) are arranged. The bottom (4) of the hollow chamber is drilled through multiple holes (5) that connect the external and internal surfaces. In general, as an example, it is preferred that each hole has a diameter ranging from 0.1 pm to 10,000 pm, preferably from 1 pm to 7,500 pm, more preferably from 10 pm to 5,500 pm and even more preferably from 100 pm to 2,500 pm. The hollow chamber (1 , 2) has a support base (3) provided with at least two inclined legs (31 , 32) with spacing between them in order to allow the flow of fluids from the culture medium (8) towards the bottom (4) with perforations (5) of the hollow chamber (1 , 2), thus reaching the hydrogel layer (6) on which the cells (7) rest. Thus, when the cell culture insert according to the present invention is positioned inside the well of a tissue culture vessel (9), the tip of each leg (31 , 32) touches the bottom edge of the said vessel (9) in order to stabilize the cell culture insert.
Note should be made of the fact that the outer dimensions of the cell culture insert object of the present invention are sufficiently smaller than the internal dimensions of the tissue culture vessel to allow the insert to be completely inserted into the tissue culture vessel. The cell culture insert according to the present invention can be sized to be adapted for use with different tissue culture vessels and vats of different sizes and geometric configurations.
The hydrogel used in the cell culture insert according to the present invention can be any of the synthetic type, such as, for example, those produced in polyethylene glycol, polyacrylic acid, polyvinyl alcohol, and polypeptides, but not limited to them. A natural hydrogel can also be used,
such as, for example, fibrin, collagen, gelatin, hydrogels derived from the decellularized extracellular matrix, hyaluronic acid, alginate, agarose, nanocellulose and modified natural polymers, but not limited to them. Mixtures of these hydrogels or compositions thereof can also be used.
Additionally, but without limitations, the hydrogel may have its surface chemically modified to present chemical functional groups, including carbonyls (C = O), alcohols (-OH), carboxylic acids (CO2H), esters (CO2R), and amines (NH2), as well as having its surface biologically modified to present specific (macro) biological molecules, including antibodies, enzymes, and hormones.
The hydrogel for use in the cell culture insert according to the present invention can also have surface topography characteristics, including columns, pits, grooves, hemispheres, and grids, among other possible ones.
The present invention also deals with a method of using the cell culture insert according to the present invention, comprising introducing target cells on top of the hydrogel (6) and a cell culture medium (8) in the vessel (9) where the insert is incorporated to grow the target cells in an air-gel interface.
The method of use according to the present invention includes chemical, biochemical, or biological compounds that are added to the medium contained in the vessel (9), so that they move from said vessel (9) to the hydrogel (6), pass through the hydrogel (6) and reach cell culture (7).
Biological agents, for example, prions, viruses, bacteria, and protozoa, without limitation, are added to the medium (8) contained in the vessel (9), so that they move from the vessel (9) to the hydrogel (6), passing through the hydrogel (6) and reaching the cell culture (7). Alternatively, or in addition, said biological agents can be added directly to the cells (7) that grow on top of the hydrogel (6), the effects of such treatments being evaluated on the target cells (7).
Those skilled in the art will appreciate that innumerable variations affecting the scope of the present invention are permitted, without thereby departing from the inventive scope disclosed herein and, thus, reinforcing the fact that the present invention is not limited to the particular configurations and embodiments described above.
Claims
1. Cell culture insert containing hydrogel, characterized by comprising a hollow chamber with two distinct portions, an upper portion (1) and a lower portion (2), and a support base (3), said upper portion (1) having an open end at the top and said lower portion (2) having side walls (21) and being closed at the bottom (4) perforated (5), provided with a hydrogel plug (6) on which are positioned the cells (7).
2. Cell culture insert containing hydrogel, according to claim 1 , characterized by the fact that the upper portion (1) of the said hollow chamber has inclined side walls (11), as the diameter of the larger upper portion (12) than the diameter of the lower portion (13).
3. Cell culture insert containing hydrogel, according to any one of claims 1 or 2, characterized by the fact that said upper portion (1) has an open upper end where one or more flaps (14) directed to the interior are provided of the hollow chamber.
4. Cell culture insert containing hydrogel, according to claim 1 , characterized by the fact that the side walls (21) are perforated are equipped with multiple holes with a diameter ranging from 0.1 pm to 10,000 pm, preferably from 1 pm to 7,500 pm, more preferably from 10 pm to 5,500 pm and even more preferably from 100 pm to 2,500 pm.
5. Cell culture insert containing hydrogel, according to claim 1 , characterized by the fact that said hydrogel plug (6) fills the complete volume of the lower portion (2) of the hollow chamber.
6. Cell culture insert containing hydrogel, according to claim 1 , characterized by the fact that said multiple holes (5) of the bottom (4) of the hollow chamber have a diameter ranging from 0.1 pm to 10,000 pm, preferably from 1 pm to 7,500 pm, more preferably from 10 pm to 5,500 pm and even more preferably from 100 pm to 2,500 pm.
7. Cell culture insert containing hydrogel, according to claim 1 , characterized by the fact that said support base (3) is provided with at least two inclined legs (31 , 32) with spacing between them.
8. Cell culture insert containing hydrogel, according to claim 1 , characterized by the fact that said inclined legs (31 , 32) are such that they touch the bottom edge of a tissue culture vessel bowl (9).
9. Cell culture insert containing hydrogel, according to any one of claims 1 or 5, characterized by the fact that the hydrogel is of the synthetic type and chosen from those produced in polyethylene glycol, polyacrylic acid, polyvinyl alcohol, and polypeptides, their mixtures or compositions containing them.
10. Cell culture insert containing hydrogel, according to any one of claims 1 or 5, characterized by
the fact that the hydrogel is of the natural type and chosen from those produced in fibrin, collagen, gelatin, hydrogels derived from decellularized extracellular matrix, hyaluronic acid, alginate, agarose, nanocellulose and modified natural polymers, their mixtures or compositions containing them.
11. Cell culture insert containing hydrogel, according to any one of claims 1 , 5, 9, or 10, characterized by the fact that the hydrogel is a mixture of the synthetic type with the natural type.
12. Cell culture insert containing hydrogel, according to any one of claims 1 , 5, 9 to 11 , characterized by the fact that the hydrogel has its surface chemically modified to present chemical functional groups, including carbonyls (C = O), alcohols (-OH), carboxylic acids (CO2H), esters (CO2R) and amines (NH2).
13. Cell culture insert containing hydrogel, according to any one of claims 1 , 5, 9 to 11 , characterized by the fact that the hydrogel has its surface biologically modified to present specific (macro) biological molecules, including antibodies, enzymes, and hormones.
14. Cell culture insert containing hydrogel, according to any one of claims 1 , 5, 9 to 13, characterized by the fact that the hydrogel has surface topography characteristics, including columns, pits, grooves, hemispheres, and grids.
15. Method of use of cell culture insert containing hydrogel as defined in any one of claims 1 to 14, characterized by the fact that it comprises: introducing the target cells on top of the hydrogel (6), introducing a cell culture medium (8) into the vessel (9), introducing chemical, biochemical, or biological compounds into the cell culture medium (8) contained in the vessel (9), allowing said chemical, biochemical, or biological compounds to move from said vessel (9) to the hydrogel (6) until the cell culture is reached (7).
16. Method of use, according to claim 15, characterized by the fact that biological compounds are chosen from prions, viruses, bacteria, and protozoa.
17. Method of use, according to any one of claims 14 to 16, characterized by the fact that chemical, biochemical or biological compounds are added directly to the cells (7) that grow on top of the hydrogel (6).
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BR102020007764-3A BR102020007764A2 (en) | 2020-04-17 | 2020-04-17 | CELL CULTURE INSERT CONTAINING HYDROGEL AND METHOD OF USE |
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WO2024068900A1 (en) * | 2022-09-29 | 2024-04-04 | Danmarks Tekniske Universitet | Cell culture insert |
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