KR101548768B1 - 3-dimensional cell chip substrate and method of co-culturing cell using the same - Google Patents

3-dimensional cell chip substrate and method of co-culturing cell using the same Download PDF

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KR101548768B1
KR101548768B1 KR1020110012957A KR20110012957A KR101548768B1 KR 101548768 B1 KR101548768 B1 KR 101548768B1 KR 1020110012957 A KR1020110012957 A KR 1020110012957A KR 20110012957 A KR20110012957 A KR 20110012957A KR 101548768 B1 KR101548768 B1 KR 101548768B1
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culture medium
cell
column
auxiliary body
hole
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KR1020110012957A
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KR20120092985A (en
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정세훈
구보성
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삼성전기주식회사
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    • 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
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C12M35/08—Chemical, biochemical or biological means, e.g. plasma jet, co-culture
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/34—Internal compartments or partitions

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Abstract

The three-dimensional cell chip substrate according to the present invention is manufactured by combining the first culture medium 200 with the well 110 of the first auxiliary body 100 to cultivate the first cells, The second cell 400 is inserted into the second hole 310 and the second cell 400 is inserted into the second auxiliary cell 100. The first cell 200 and the second cell 400 are connected to the first cell 100, And the second auxiliary body 200, and are bonded to each other, so that the xenogeneic cells can be easily co-cultured. In addition, in a state where the first culture medium 200 and the second culture medium 400 are combined, the first cells surround the second cells, and interaction among the cells is active.

Description

The present invention relates to a three-dimensional cell chip substrate and a co-culturing method using the same,

The present invention relates to a cell co-culture method using a three-dimensional cell chip substrate.

The process of developing new reagents (or drugs) is complex and requires cell culture to test the efficacy and toxicity of candidate reagents. In general, the cell culture method is a 2D cell monolayer culture in which cells are adhered to a two-dimensional surface, a cell is fixed three-dimensionally in a three-dimensional biomatrix, or a spheroid And 3D cell culture, which is a method of cultivation.

The two-dimensional cell chip is a microtiter plate (for example, 6-well, 12-well, 24-well, 96-well, 384-well, 1536- Microtiter plates, etc.). The culture medium required for culturing the cells in the wells of such a microtiter plate is about several ml to several tens of micro liter. These microtiter plates have the advantage of being able to perform a variety of simple experiments quickly at low cost compared to animal / human clinical trials.

However, since the microtiter plate has a form in which cells are immobilized in a well, there is a problem in a process for recovering cells after treating the reagent. This problem becomes even more pronounced when the size of the well is reduced and the number of wells is increased in order to perform more experiments on one plate, such as a 384-well microtiter plate or a 1536-well microtiter plate.

Solidus Biosciences has developed an array-based three-dimensional cell chip that cultivates three-dimensionally immobilized cells on a flat glass substrate. The three-dimensional cell chip does not form a well, but uses cells such as collagen, alginate, or matrigel to fix the cells on a glass substrate. The three-dimensional cell chip is similar to the actual biomedical structure as the two-dimensional cell chip, and has an advantage of matching with the in-vivo result.

However, in the conventional three-dimensional cell chip, there is a problem that it is difficult to bond and separate the cell chips when co-culturing the xenogeneic cells. Cell co-cultivation for cell interaction studies is crucial for cell growth, migration, and differentiation, and is essential for therapeutic studies in which tissue functions are performed by intercellular interactions. Therefore, there is a need to develop a cell chip substrate capable of three-dimensional cell culture as well as co-culturing of xenogeneic cells.

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a three-dimensional cell chip substrate capable of culturing three-dimensional cells and facilitating cell co-culture.

A three-dimensional cell chip substrate according to a preferred embodiment of the present invention includes a first auxiliary body including a well formed at the center and a column protruding from the center of the well, a second auxiliary body including a second hole formed at the center, and a second auxiliary body having a column corresponding to the second hole of the second auxiliary body, wherein the first hole corresponds to the column, 2 culture medium.

Here, the present invention is characterized in that the first cell is cultured on the upper surface of the first culture medium, and the second cell is cultured on the upper surface of the column of the second culture medium.

Further, the present invention is characterized in that the shape and diameter of the first hole of the first culture medium are the same as the shape and diameter of the cross section of the column of the first auxiliary body.

Further, the present invention is characterized in that the shape and diameter of the cross section of the column of the second culture medium are the same as the shape and diameter of the second hole of the second auxiliary body.

Further, the present invention is characterized in that the shape and diameter of the first hole of the first culture medium are the same as the shape and diameter of the cross section of the column of the second culture medium.

Further, the present invention is characterized in that the thickness of the first culture medium is smaller than the depth of the well of the first auxiliary body.

In addition, the present invention is characterized in that the height of the column of the second culture medium is smaller than the thickness of the second auxiliary body.

Further, the present invention is characterized in that the thickness of the first culture medium and the height of the column of the second culture medium are the same.

In addition, the present invention is characterized in that a first matrix including a first cell is formed on an upper surface of the first culture medium, and a second matrix including a second cell is formed on an upper surface of the column of the second culture medium.

Further, the present invention is characterized by further comprising a bar-shaped handle formed on the upper surface of the first culture medium and the second auxiliary body.

A cell co-culturing method according to a preferred embodiment of the present invention comprises the steps of (A) culturing a first auxiliary body including a well formed at the center and a column protruding to the center of the well, A step of culturing the first cell on the upper surface of the first culture medium by binding a first culture medium on which the first hole is formed, (B) forming a column on the second hole of the second auxiliary body including the second hole formed in the center Culturing the first cell and the second culture medium on the upper surface of the column of the second auxiliary body by inserting the column of the second culture medium, (C) Separating the first cell and the second cell from the second culture medium; and (D) co-culturing the first cell and the second cell by inserting the column of the second culture medium into the first hole of the first culture medium .

Here, the present invention is characterized in that the shape and diameter of the first hole of the first culture medium are the same as the shape and diameter of the cross section of the column of the first auxiliary body.

Further, the present invention is characterized in that the shape and diameter of the cross section of the column of the second culture medium are the same as the shape and diameter of the second hole of the second auxiliary body.

Further, the present invention is characterized in that the shape and diameter of the first hole of the first culture medium are the same as the shape and diameter of the cross section of the column of the second culture medium.

Further, the present invention is characterized in that the thickness of the first culture medium is smaller than the depth of the first auxiliary chuck well.

In addition, the present invention is characterized in that the height of the column of the second culture medium is smaller than the thickness of the second auxiliary body.

Further, the present invention is characterized in that the thickness of the first culture medium and the height of the column of the second culture medium are the same.

In addition, the present invention is characterized in that a first matrix including a first cell is formed on an upper surface of the first culture medium, and a second matrix including a second cell is formed on an upper surface of the column of the second culture medium.

Further, the present invention is characterized in that the present invention further comprises a rod-like handle formed on the upper surface of the first culture medium, the first auxiliary body, the second culture medium, and the second auxiliary body.

Prior to that, terms and words used in the present specification and claims should not be construed in a conventional and dictionary sense, and the inventor may properly define the concept of the term in order to best explain its invention It should be construed as meaning and concept consistent with the technical idea of the present invention.

According to the present invention, a first cell is cultured by combining a first culture medium and a first auxiliary body, a second culture medium is cultured by binding a second culture medium and a second auxiliary body, and then a first culture medium and a second culture medium It is possible to easily co-culture differentiated cells by separating the first and second auxiliary bodies from each other and combining the first culture medium and the second culture medium with each other.

1A and 1B are a perspective view and a cross-sectional view of a first auxiliary body according to a preferred embodiment of the present invention.
2A and 2B are a perspective view and a cross-sectional view of a first culture according to a preferred embodiment of the present invention.
3A and 3B are a perspective view and a cross-sectional view of a second auxiliary body according to a preferred embodiment of the present invention.
4A and 4B are a perspective view and a cross-sectional view of a second culture medium according to a preferred embodiment of the present invention.
FIGS. 5 to 12 are a perspective view and a cross-sectional view, respectively, of a cell co-culturing method according to a preferred embodiment of the present invention.
FIG. 13 is a plan view showing a coupling structure of a first culture medium and a second culture medium according to the present invention bonded to a well plate. FIG.

BRIEF DESCRIPTION OF THE DRAWINGS The objectives, specific advantages and novel features of the present invention will become more apparent from the following detailed description taken in conjunction with the accompanying drawings, in which: FIG. It should be noted that, in the present specification, the reference numerals are added to the constituent elements of the drawings, and the same constituent elements are assigned the same number as much as possible even if they are displayed on different drawings. In the following description, well-known functions or constructions are not described in detail since they would obscure the invention in unnecessary detail.

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.

A three-dimensional cell chip substrate according to the present invention comprises a first auxiliary body having a column at the center of a well and a well, a first culture medium in which a first cell is cultured on a top surface thereof, A second auxiliary body having a hole formed therein, and a second culture medium having a column on which an upper surface of the second cell is cultured.

The present invention relates to a method for culturing a first cell, which comprises culturing a first cell by culturing a first cell and culturing the second cell by inserting a column of the second culture into a second hole of the second cell, , The first culture medium and the second culture medium are separated from the first and second auxiliary bodies, respectively, and are bonded to each other, so that the xenogeneic cells can be easily co-cultured. Hereinafter, the components of the three-dimensional cell chip substrate according to the present invention will be described in detail.

First, the first auxiliary body 100 includes a well 110 formed at the center as shown in FIGS. 1A and 1B and a column 130 formed at the center of the well 110. The first auxiliary body 100 is used as an auxiliary application for culturing the first cells in the first culture medium 200 by binding the first culture medium 200 to the well 110. [ Although the first auxiliary body 100 having the well 110 formed in the shape of the cylinder 130 is shown in FIG. 1A, the present invention is not limited to this. Lt; / RTI > It is preferable that the diameter Da 1 of the first auxiliary member 100 is 15 mm to 25 mm and the diameter Da 2 of the cross section of the column 130 is 6 mm to 10 mm.

At this time, the first auxiliary body 100 may be made of glass, plastic, ceramics, or the like. However, the present invention is not limited to this, and any material having sufficient rigidity to support the first culture medium 200 coupled to the well 110 and having high biocompatibility may be used.

In the present invention, the term 'cell' includes various nucleic acid molecules (for example, DNA, RNA, oligonucleotide, cDNA, plasmid and the like), peptide peptides, proteins, lipids, proteins or lipid membranes, organic or inorganic chemical molecules (e.g., pharmaceuticals or compounds elsewhere), virus particles, eukaryotic or prokaryotic cells, .

Next, as shown in FIGS. 2A and 2B, the first culture medium 200 has a first hole 210 formed at the center thereof. The first culture medium 200 is combined with the first auxiliary body 100 to cultivate the first cells on the upper surface thereof. Specifically, the column 130 of the first auxiliary body 100 is inserted into the first hole 210 of the first culture vessel 200, and the first culture vessel 200 is connected to the well 110 Then, the first cells are fixed on the upper surface of the first culture medium 200 and cultured in an incubator. The first culture medium 200 in which the first cells are cultured on the upper surface may be co-cultured with the second culture medium 400 in which the second cells are cultured after being separated from the first auxiliary body 100 .

The shape and diameter Db 2 of the first hole 210 of the first culture medium 200 are preferably equal to the shape and diameter Da 2 of the cross section of the column 130 of the first auxiliary body 100 Do. In this case, there is no space between the first hole 210 and the column 130 in the state where the column 130 of the first auxiliary body 100 is inserted into the first hole 210, Can be supplied to the first cells without spillage.

The thickness Tb 1 of the first culture medium 200 is smaller than the depth Ha 1 of the well 110 of the first auxiliary body 100. The depth Ha 1 of the well 110 of the first assistant body 100 and the depth Ha 1 of the first auxiliary body 100 in the state where the first culture medium 200 is coupled to the well 110 of the first auxiliary body 100, The culture space having the height of the difference of the thickness (Tb 1 ) of the culture medium 200 is formed on the upper surface of the first culture medium 200, whereby the three-dimensional cell culture can be performed.

Here, the term "three-dimensional cell culture" refers to a structure in which cells are contained in a matrix and have a three-dimensional structure similar to an actual living body structure and cultured so that intercellular interactions occur three-dimensionally. Two-dimensional cell cultures performed in conventional two-dimensional impermeable planes are unable to accurately simulate the cellular environmental conditions and thus limitations exist in insights into the mechanisms of cell movement and transmission and disease. The three-dimensional cell culture is performed under culture conditions similar to the actual bio-structure, thereby providing new insights and opportunities for drug inspections for the analysis and treatment of diseases. In addition, three-dimensional cell culture can be used to form a tissue similar to the cellular environment in the human body, and a three-dimensional microstructure can be realized through natural interaction between cells and cells.

The matrix fixes the cells three-dimensionally inside and provides an environment similar to the biotissue, and stores the drug and the culture solution to supply the cells. The matrix material may be a sol-gel, an inorganic material, an organic polymer, or an organic-inorganic composite material. In particular, the matrix may be a hydrogel having a porous structure and having no toxicity to the biomaterial, such as an extracellular matrix, such as collagen in which the fluid moves through diffusion, alginate or matrigel, Is preferably employed. The present invention is not only capable of forming a matrix 250 (see FIG. 6) containing a first cell on the top surface of a first culture medium 200 to perform three-dimensional cell culture, but also simply forming a single layer of a first cell Two-dimensional cell culture can also be performed.

Although FIG. 2A illustrates a first culture medium 200 having a first hole 210 formed in a cylindrical shape, the first culture medium 200 is not limited to the above-described shape, and may have various shapes such as a bar shape, a triangular column, . However, the shape of the first culture medium 200 should correspond to the shape of the well 110 so that the first culture medium 200 can be coupled to the well 110 of the first auxiliary body 100. It is preferable that the diameter Db 1 of the first culture medium 200 is 11 mm to 17 mm and the diameter Db 2 of the first hole 210 is 6 mm to 10 mm.

In addition, the first culture medium 200 may further include a rod-shaped handle 230 formed on an upper surface of the upper surface of the first culture medium 200. By forming the handle 230, the first culture medium 200 can be easily combined and separated.

Next, the second auxiliary body 300 includes a second hole 310 formed at the center as shown in FIGS. 3A and 3B. The second auxiliary body 300 is used as an auxiliary application for culturing the second cell in the second culture medium 400 by inserting the column 410 of the second culture medium 400 into the second hole 310 .

Although the second auxiliary body 300 having the second hole 310 formed in a cylindrical shape is shown in FIG. 3A, the second auxiliary body 300 is not limited to the above-described shape, and may have various shapes such as a bar shape, have. It is preferable that the diameter Dc 1 of the second auxiliary body 300 is 11 mm to 17 mm and the diameter Dc 2 of the second hole 310 is 6 mm to 10 mm.

In addition, the second auxiliary body 300 may further include a bar-shaped handle 330 formed on an upper surface of the upper body. By forming the handle 330, the second auxiliary body 300 can be easily engaged and separated.

Next, the second culture medium 400 includes a column 410 corresponding to the second hole 310 as shown in FIGS. 4A and 4B. The column 410 of the second culture medium 400 is inserted into the second hole 310 of the second auxiliary body 300 and the second cell is cultured on the upper surface of the column 410.

At this time, the shape and diameter Dd 2 of the cross section of the column 410 of the second culture medium 400 are preferably the same as the shape and diameter Dc 2 of the second hole of the second auxiliary body 300. A space between the second hole 310 and the column 410 may be formed in a state where the column 410 of the second culture medium 400 is inserted into the second hole 310 of the second auxiliary body 300 So that the reagent or the culture liquid can be supplied to the second cell without spillage.

The height Hd 1 of the column 410 of the second culture medium 400 is smaller than the thickness Tc 1 of the second auxiliary body 300. The height Hd 1 of the column 410 of the second culture medium 400 and the thickness Tc 1 of the second auxiliary body 300 are set so that the second culture medium 400 is coupled to the second auxiliary body 300, A culture space having a height equal to the height of the second culture medium 400 is formed on the upper surface of the column 410 of the second culture medium 400 to coat the upper surface of the column 410 with a matrix 430 The culture can be performed. As described above, a single layer of the second cell may be formed to perform two-dimensional cell culture.

4A shows a second culture medium 400 having a column 410 formed on a part of a top surface of a cylinder. However, the shape of the second culture medium 400 is not limited to the above-described shape, and may be various shapes such as a bar shape or a triangular column. However, the column 410 of the second culture medium 400 to be inserted into the second hole 310 of the second auxiliary body 300 should have a shape corresponding to the second hole 310. The diameter Dd 1 of the second culture medium 400 is 11 mm to 17 mm and the diameter Dd 2 of the cross section of the column 410 is preferably 6 mm to 10 mm.

After the cells are cultured, the first culture medium 200 and the second culture medium 400 are separated from the first auxiliary body 100 and the second auxiliary body 300, respectively, The shape and diameter (Db 2 ) of the first hole 210 of the first culture medium 200 are set to be the same as the diameter of the first hole 210 of the first culture medium 200 because the cells 210 of the second culture medium 400 are inserted into the hole 210, Is preferably the same as the shape and diameter (Dd 2 ) of the cross section of the column 410 of the second culture medium 400. In a state where the first culture medium 200 and the second culture medium 400 are combined, the first cell and the second cell are in contact with each other, and interaction between cells occurs. Here, intercellular interactions include intercellular signaling through body fluids such as growth hormone, cytokine, etc. and signaling by direct contact between cells. In addition, cancer-associated fibroblast (CAF) also includes indirect effects such as enhancing the viability of cancer cells by creating an environment in which cancer cells are diffused.

At this time, it is preferable that the thickness Tb 1 of the first culture medium 200 and the height Hd 1 of the column 410 of the second culture medium 400 are the same. In this case, the upper surface of the first culture medium 200 and the upper surface of the column 410 of the second culture medium 400 are positioned on the same plane with the second culture medium 400 coupled to the first culture medium 200 , The contact area between the first cell and the second cell is maximized.

The method for co-culturing cells according to a preferred embodiment of the present invention comprises the steps of (A) combining a first culture medium 200 with a well 110 of a first auxiliary medium 100, (B) inserting the column 410 of the second culture medium 400 into the second hole 310 of the second auxiliary body 300 to form the column 410 of the second culture medium 400, (C) separating the first culture medium 200 and the second culture medium 400 from the first auxiliary body 100 and the second auxiliary body 300, respectively, and D) Inserting the column 410 of the second culture medium 400 into the first hole 210 of the first culture medium 200 co-cultivates the first cell and the second cell. The cell co-culturing method according to the present invention is advantageous in that the structure for co-culturing xenogeneic cells can be easily joined and separated. The second cell cultured on the upper surface of the column 410 of the second culture cell 400 may be cultured on the upper surface of the first culture cell 200 in a state where the first culture medium 200 and the second culture medium 400 are combined, The first cell is enclosed and the interaction of the xenogeneic cells becomes active. Hereinafter, the cell co-culture method will be described in order. I will omit or duplicate the previous sections.

5 and 6, a first culture 200 is coupled to a well 110 of a first auxiliary body 100 to form a first cell 200 on an upper surface of the first culture 200 Lt; / RTI > Specifically, the column 130 of the first auxiliary body 100 is inserted into the first hole 210 of the first culture medium 200, and the first culture medium 200 is inserted into the well 200 of the first auxiliary body 100, (110) well. Next, the matrix 250 containing the first cells may be coated on the upper surface of the first culture medium 200 to perform three-dimensional cell culture. As a method of coating the matrix 250 on the upper surface of the first culture medium 200, spotting, contact printing, piezoelectric printing, micro-pipetting, and the like are available. Alternatively, the first cell may be coated on the upper surface of the first culture medium 200 as a single layer to perform two-dimensional cell culture.

At this time, the upper surface of the first culture medium 200 may be chemically surface-treated so as to have a functional group before culturing the first cells. Since the upper surface of the first culture medium 200 has a functional group, the matrix 250 containing the first cells can be easily fixed. Examples of the functional group include an amino group, an aldehyde group, an epoxide group and an ester group. The chemical surface treatment may be performed by spin coating, dip coating, roll coating, spray coating, or the like on the upper surface of the first culture medium 200 with a solution containing a functional group .

In addition, the upper surface of the first culture medium 200 may be surface-treated with hydrophobicity before culturing the first cells. When the upper surface of the first culture medium 200 is subjected to the hydrophobic surface treatment, the matrix 250 containing the first cells may not spread, and a uniform shape may be maintained. The hydrophobic surface treatment may be performed by performing MVD (molecular vapor deposition), PVD (physical vapor deposition), CVD (chemical vapor deposition), or the like on the upper surface of the first culture medium 200.

Next, as shown in FIGS. 7 and 8, the column 410 of the second culture medium 400 is inserted into the second hole 310 of the second auxiliary body 300, The second cell is cultured on the upper surface of the cell 410. As described above, the matrix 430 containing the second cells may be coated on the top surface of the column 410 of the second culture medium 400 to perform three-dimensional cell culture. In addition, two-dimensional cell culture can be performed by coating the second cell on the upper surface of the column 410 of the second culture medium 400 with a single layer.

In this case, the upper surface of the column 410 of the second culture medium 400 may be chemically surface-treated or hydrophobically treated so as to have a functional group, like the upper surface of the first culture medium 200, before culturing the second cells.

Next, as shown in FIGS. 9 and 10, the first culture medium 200 and the second culture medium 400 are separated from the first and second auxiliary bodies 100 and 300, respectively. The matrices 250 and 430 formed on the upper surfaces of the columns 410 and 410 of the first culture medium 200 and the second culture medium 400 are in a gel state to separate the first and second auxiliary bodies 100 and 300, (250, 430) do not flow down and have a fixed shape.

Next, as shown in FIGS. 11 and 12, the column 410 of the second culture medium 400 is inserted into the first hole 210 of the first culture medium 200, Lt; / RTI > When the first culture medium 200 and the second culture medium 400 are combined, the matrix 250 including the first cells is surrounded by the matrix 430 including the second cells as shown in FIG. 12, Action.

In addition, as shown in FIG. 13, the binding structure of the first culture medium 200 and the second culture medium 400 may be inserted into a well plate 500 to co-culture the cells. The present invention is advantageous in that it is excellent in compatibility with a well plate (500) used as a conventional cell culture medium, and is convenient to use. In addition, by inserting a plurality of binding structures of the first culture medium 200 and the second culture medium 400 into a well plate 500, various drug tests can be simultaneously performed.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it should be understood that the present invention is not limited to the above-described three-dimensional cell chip substrate and method for co- It will be apparent that modifications and improvements may be made by those skilled in the art without departing from the spirit and scope of the invention. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

100: first auxiliary body 110: well,
130, 410: column 200: first culture medium
210: first hole 230, 330: handle
250, 430: matrix 300: second auxiliary body
310: second hole 400: second culture medium
500: well plate

Claims (19)

A first hole corresponding to the column is formed to be coupled to the well of a first auxiliary body including a well formed at the center and a column protruding from the center of the well, 1 cells are cultured and separated from the first auxiliary body; And
A column corresponding to the second hole is formed so as to be coupled to the second hole of a second auxiliary body including a second hole formed in the center, a second cell is cultured on the upper surface of the column, A separated second culture; Lt; / RTI >
And a column of a second culture medium separated from the second auxiliary body is inserted into a first hole of a first culture medium separated from the first auxiliary body. delete The method according to claim 1,
Wherein a shape and a diameter of the first hole of the first culture medium are the same as a shape and a diameter of a cross section of the column of the first auxiliary body. The method according to claim 1,
And the shape and diameter of the cross section of the column of the second culture medium are the same as the shape and diameter of the second hole of the second auxiliary body. The method according to claim 1,
Wherein a shape and a diameter of the first hole of the first culture medium are the same as a shape and a diameter of a cross section of the column of the second culture medium. The method according to claim 1,
Wherein the thickness of the first culture medium is smaller than the depth of the well of the first auxiliary body. The method according to claim 1,
And the height of the column of the second culture medium is smaller than the thickness of the second auxiliary body. The method according to claim 1,
Wherein the thickness of the first culture medium and the height of the column of the second culture medium are the same. The method according to claim 1,
Wherein a first matrix including a first cell is formed on an upper surface of the first culture medium and a second matrix including a second cell is formed on an upper surface of the column of the second culture medium. The method according to claim 1,
Further comprising a rod-like handle formed on an upper surface of the first culture medium and the second auxiliary body. (A) a first culture medium having a first hole formed in the well of a first auxiliary body including a well formed at the center and a column protruding from the center of the well, Culturing the first cell on the upper surface of the culture medium;
(B) inserting the column of the second culture medium in which the column is formed in the second hole of the second auxiliary body including the second hole formed at the center, culturing the second cell on the upper surface of the column of the second auxiliary body ;
(C) separating the first culture medium and the second culture medium from the first auxiliary body and the second auxiliary body, respectively; And
(D) co-culturing the first cell and the second cell by inserting the column of the second culture medium into the first hole of the first culture medium;
/ RTI > The method of claim 1, The method of claim 11,
Wherein the shape and diameter of the first hole of the first culture medium are the same as the shape and diameter of the cross section of the column of the first auxiliary body. The method of claim 11,
And the shape and diameter of the cross section of the column of the second culture medium are the same as the shape and diameter of the second hole of the second auxiliary body. The method of claim 11,
Wherein the shape and diameter of the first hole of the first culture medium are the same as the shape and diameter of the cross section of the column of the second culture medium. The method of claim 11,
Wherein the thickness of the first culture medium is smaller than the depth of the well of the first auxiliary body. The method of claim 11,
And the height of the column of the second culture medium is smaller than the thickness of the second auxiliary body. The method of claim 11,
Wherein the thickness of the first culture medium and the height of the column of the second culture medium are the same. The method of claim 11,
Wherein a first matrix including a first cell is formed on an upper surface of the first culture medium and a second matrix including a second cell is formed on an upper surface of the column of the second culture medium. The method of claim 11,
Further comprising a rod-shaped handle formed on an upper surface of the first culture medium, the first auxiliary body, the second culture medium, and the second auxiliary body.
KR1020110012957A 2011-02-14 2011-02-14 3-dimensional cell chip substrate and method of co-culturing cell using the same KR101548768B1 (en)

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US20040214313A1 (en) 2003-04-28 2004-10-28 Weihua Zhang Cell interaction culture system and uses thereof
KR100718158B1 (en) 2006-04-21 2007-05-14 삼성전자주식회사 Cell co-culture device
JP2007215472A (en) 2006-02-16 2007-08-30 Olympus Corp Cell culture container

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US20040214313A1 (en) 2003-04-28 2004-10-28 Weihua Zhang Cell interaction culture system and uses thereof
JP2007215472A (en) 2006-02-16 2007-08-30 Olympus Corp Cell culture container
KR100718158B1 (en) 2006-04-21 2007-05-14 삼성전자주식회사 Cell co-culture device

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