# US 2021/0340475 A1 · Apparatus for cell culture and method for cell culture using the same

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- Number: US 2021/0340475 A1
- Document ID: US20210340475A1
- Status: Published application
- Kind: A1
- Application number: 17/238,332
- Filed: 2021-04-23
- Published: 2021-11-04
- Earliest claimed: 2020-04-29
- Assignees: Postech Research and Business Development Foundation; Board of Regents, The University of Texas System
- Applicants: POSTECH Research and Business Development Foundation; Board of Regents, The University of Texas System
- Inventors: Dong Sung KIM; Woojung Shin; Hyun Jung Kim; Jae Seung Yoon; Hyeon Beom Chong
- Web page: https://patentgrep.com/patent/US20210340475A1

## CPC

- C12M 23/12: {Well or multiwell plates (C12M25/04 takes precedence)}
- C12M 23/24: {Gas permeable parts}
- C12M 25/04: {in combination with well or multiwell plates, i.e. culture inserts}
- C12M 29/04: {Filters; Permeable or porous membranes or plates, e.g. dialysis}
- C12M 41/32: {of substances in solution}
- C12N 5/0068: {General culture methods using substrates (for specific animal cell type C12N5/06)}
- C12N 5/06: {Animal cells or tissues; Human cells or tissues (preservation of excised living parts A01N1/10)}
- 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}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present disclosure provides a three-dimensional cell culture method and a cell culture apparatus using the same. The cell culture method includes lowering, when culturing cells on at least one surface of a porous support, a concentration of a cell-secreted substance in a lower region of the other surface of the porous support. The cell culture apparatus includes: an upper chamber including an opening, porous support having at least one surface on which cells are cultured, and a cell culture space in which a culture medium is housed; a lower chamber including a space in which at least one upper chamber is disposed; and a device lowering a concentration of a cell-secreted substance in a lower region of the other surface of the porous support.

## Claims

### Claim 1

A cell culture method comprising lowering a concentration of a cell-secreted substance in a lower region of the other surface of the porous support, when culturing cells on at least one surface of a porous support.

### Claim 2 (depends on 1)

The cell culture method of claim 1, wherein the lower region of the other surface of the porous support includes a region spaced apart from the porous support by 0.01 mm.

### Claim 3 (depends on 1)

The cell culture method of claim 1, wherein in the lowering of the concentration of the cell-secreted substance, the concentration of the cell-secreted substance is maintained at 100 μg/ml or less.

### Claim 4 (depends on 1)

The cell culture method of claim 1, wherein the cell-secreted substance includes at least one component selected from the group consisting of sFRP1, FRZB, Sizzled, Sizzled2, Crescent, WIF-1, Cerberus, Coco, DKK-2, DKK-3, DKK-4, Soggy, sFRP2, sFRP3, sFRP4, sFRP5, and DKK-1.

### Claim 5 (depends on 1)

The cell culture method of claim 1, wherein in the lowering of the concentration of the cell-secreted substance, a concentration of DKK-1 is maintained at 10 μg/ml or less.

### Claim 6 (depends on 1)

The cell culture method of claim 1, wherein the lowering of the concentration of the cell-secreted substance in the lower region is performed by i) supplying a flowing fluid to the lower region, ii) applying a motion of fluid to the lower region, or iii) adding an antibody against the cell-secreted substance.

### Claim 7 (depends on 6)

The cell culture method of claim 6, wherein the motion of the fluid is applied by at least one selected from the group consisting of a reciprocating motion of a fluid, a rotational motion of a fluid, vortex formation of a fluid, a gradient motion of a fluid, and a random motion of a fluid.

### Claim 8 (depends on 1)

The cell culture method of claim 1, wherein the cell is an epithelial cell.

### Claim 9 (depends on 1)

The cell culture method of claim 1, wherein the cell is an intestinal epithelial cell.

### Claim 10

A cell culture apparatus comprising:
an upper chamber including an opening, a porous support having at least one surface on which cells are cultured, and a cell culture space in which a culture medium is housed;
a lower chamber including a space in which at least one upper chamber is disposed; and
a device lowering a concentration of a cell-secreted substance in a lower region of the other surface of the porous support.

### Claim 11 (depends on 10)

The cell culture apparatus of claim 10, wherein the lower region of the other surface of the porous support includes a region spaced apart from the porous support by 0.01 mm.

### Claim 12 (depends on 10)

The cell culture apparatus of claim 10, wherein the device lowering the concentration of the cell-secreted substance in the lower region of the other surface of the porous support is i) a device supplying a flowing fluid to the lower chamber or ii) a device applying a motion of fluid to the lower chamber.

### Claim 13 (depends on 12)

The cell culture apparatus of claim 12, wherein the device applying the motion of the fluid is at least one device selected from the group consisting of a pump, a shaker, an agitator, and an ultrasonic device.

### Claim 14 (depends on 12)

The cell culture apparatus of claim 12, wherein the device supplying the flowing fluid to the lower chamber includes:
a fluid inlet and a fluid outlet provided in the lower chamber; and
a device inducing a flow of the fluid from the fluid inlet to the fluid outlet.

### Claim 15 (depends on 10)

The cell culture apparatus of claim 10, further comprising a plate through which at least one upper chamber penetrates to be inserted and fixed thereto.

### Claim 16 (depends on 10)

The cell culture apparatus of claim 10, wherein the porous support includes a nanofiber network having nanoscale pores, microscale pores, and a combination thereof, a porous thin membrane, or a combination thereof.