# US 2020/0399571 A1 · Cell culturing device and method

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- Number: US 2020/0399571 A1
- Document ID: US20200399571A1
- Status: Published application
- Kind: A1
- Application number: 16/935,250
- Filed: 2020-07-22
- Published: 2020-12-24
- Earliest claimed: 2018-01-23
- Assignees: Bar-Ilan University
- Applicants: Bar-Ilan University
- Inventors: Mordechai Deutsch; Naomi Zurgil; Elena Afrimzon; Sergei Moshkov; Orit Ravid-Hermesh; Yana Shafran; Maria Sobolev
- Web page: https://patentgrep.com/patent/US20200399571A1

## CPC

- C12M 23/12: {Well or multiwell plates (C12M25/04 takes precedence)}
- C12M 23/20: {Material Coatings (immunocoatings C12M25/00)}
- C12M 23/22: {Transparent or translucent parts (glassware for laboratory use B01L3/00)}
- C12M 25/04: {in combination with well or multiwell plates, i.e. culture inserts}
- C12M 25/06: {Plates; Walls; Drawers; Multilayer plates}
- C12M 25/14: {Scaffolds; Matrices (in general C12N5/0068)}
- C12N 5/0062: {General methods for three-dimensional culture}
- C12N 5/0075: {using microcarriers}
- C12N 2513/00: 3D culture
- 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

A cell culturing device and method of using same are provided. Embodiments of the cell culturing device include a plate having at least one well with a through-hole formed at a bottom wall thereof and a hydrogel matrix disposed in the through hole. The cell culturing device can also include an optically transparent plate at the bottom of the through-hole.

## Claims

### Claim 1

A cell culturing device comprising:
a) a plate having at least one well with a through-hole formed at a bottom wall thereof; and
b) a hydrogel matrix disposed in said through hole.

### Claim 2 (depends on 1)

The device of claim 1, further comprising:
c) at least one chamber formed in said hydrogel matrix.

### Claim 3 (depends on 1)

The device of claim 1, wherein said through-hole is shaped so as to trap said hydrogel matrix therewithin.

### Claim 4 (depends on 1)

The device of claim 1, wherein said hydrogel matrix extends into said at least one well.

### Claim 5 (depends on 1)

The device of claim 1, further comprising an optically transparent support positioned under said plate, wherein said hydrogel matrix disposed in said through-hole contacts a top surface of said support.

### Claim 6 (depends on 1)

The device of claim 1, wherein said through hole is shaped as a truncated cone.

### Claim 7 (depends on 3)

The device of claim 3, wherein an inner surface of said through hole includes at least one undercut region.

### Claim 8 (depends on 3)

The device of claim 3, wherein an inner surface of said through hole includes protrusions directed radially inward.

### Claim 9 (depends on 2)

The device of claim 2, comprising a plurality of picoliter to microliter chambers formed in said hydrogel matrix.

### Claim 10 (depends on 1)

The device of claim 1, further comprising a ring positionable in said at least one well, said ring including a circumferential inner groove.

### Claim 11 (depends on 1)

The device of claim 1, further comprising a double ring insert positionable in said at least one well, said double ring insert including a central opening defined by an inner ring of said double ring insert and a plurality of compartments defined between said inner ring and an outer ring of said double ring insert.

### Claim 12

A method of manufacturing a culturing device comprising:
a) providing a plate having at least one well with a through-hole formed at a bottom wall thereof;
b) filling said through-hole with a hydrogel; and
c) embossing at least one cell culturing chamber in said hydrogel.

### Claim 13 (depends on 12)

The method of claim 12, further comprising positioning a double ring insert within said well prior to (b).

### Claim 14 (depends on 13)

The method of claim 13, wherein said double ring insert includes a central opening defined by an inner ring of said double ring insert and a plurality of compartments defined between said inner ring and an outer ring of said double ring insert.

### Claim 15 (depends on 14)

The method of claim 14, wherein said double ring insert includes at least one circumferential groove within an inner wall of said inner ring for trapping said hydrogel.

### Claim 16

A cell culturing device comprising:
a) a plate having at least one well with a through-hole formed at a bottom wall thereof;
b) a hydrogel matrix disposed in said through-hole; and
c) a gel disposed on top of said hydrogel matrix.

### Claim 17 (depends on 16)

The device of claim 16, wherein said gel includes at least one extracellular matrix (ECM) component.

### Claim 18

(canceled)

### Claim 19 (depends on 16)

The device of claim 16, wherein said gel including at least one extracellular matrix (ECM) component is disposed within a double ring insert positioned in said at least one well, said double ring insert including a central opening defined by an inner ring of said double ring insert and a plurality of compartments defined between said inner ring and an outer ring of said double ring insert.

### Claim 20

(canceled)

### Claim 21 (depends on 9)

A method of culturing one or more cell types comprising:
(a) providing the cell culturing device of claim 9;
(b) seeding one or more cell types within said picoliter to microliter chamber; and
(c) subjecting the cell culturing device to conditions suitable for culturing said one or more cell types.

### Claim 22 (depends on 21)

The method of claim 21, wherein said chamber is formed within a plurality of compartments each being for seeding a cell type.