# US 2019/0169566 A1 · Bioreactor system

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- Number: US 2019/0169566 A1
- Document ID: US20190169566A1
- Status: Published application
- Kind: A1
- Application number: 16/269,550
- Filed: 2019-02-06
- Published: 2019-06-06
- Earliest claimed: 2010-08-16
- Assignees: Kiyatec Inc.
- Applicants: Kiyatec Inc.
- Inventors: Matthew R. GEVAERT; David E. ORR; Margaret Rebecca WIDENER
- Web page: https://patentgrep.com/patent/US20190169566A1

## CPC

- C12N 5/0062: {General methods for three-dimensional culture}
- C12M 21/08: {for producing artificial tissue or for ex-vivo cultivation of tissue (prostheses A61F2/00, grafts A61L27/00)}
- C12M 23/44: {Multiple separable units; Modules}
- C12M 25/14: {Scaffolds; Matrices (in general C12N5/0068)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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}

## Abstract

Disclosed herein are bioreactor systems and methods of utilizing said systems.

## Claims

### Claim 1

A bioreactor system for 3D passaging of cells, comprising:
at least one cell module defining a first cell culture chamber, an inlet, an outlet, and a port opening, wherein the port opening is on one end of the cell culture chamber; and
a material composition comprising two or more materials, wherein the two or more materials comprises a stiff culture material and a soft culture material that is introduced thereon to the stiff culture material, and wherein the material composition is loaded therein the at least one cell module.

### Claim 2 (depends on 1)

The bioreactor system of claim 1, wherein the stiff culture material has a porosity between 50 μm and 2 mm.

### Claim 3 (depends on 1)

The bioreactor system of claim 1, wherein the stiff culture material is made from metal, ceramic, glass, or a synthetic polymer.

### Claim 4 (depends on 1)

The bioreactor system of claim 1, wherein the soft culture material comprises a polymer of biological origin, a synthetic polymer, or both.

### Claim 5 (depends on 4)

The bioreactor system of claim 4, wherein the soft culture material is a hydrogel or uncrosslinked oligomers of polymers.

### Claim 6 (depends on 5)

The bioreactor system of claim 5, further comprising a cell encapsulated in the hydrogel.

### Claim 7 (depends on 1)

The bioreactor system of claim 1, wherein the soft culture material releases from the stiff culture material by chemical degradation, light exposure, change in temperature, change in pH, or exposure to a chemical catalyst.

### Claim 8 (depends on 1)

The bioreactor system of claim 1, wherein the stiff culture material is composed of beads or microparticles.

### Claim 9 (depends on 1)

The bioreactor system of claim 1, wherein the stiff culture material comprises a porous matrix.

### Claim 10 (depends on 1)

The bioreactor system of claim 1, wherein the soft culture material is introduced into or onto the stiff culture material prior to being loaded into the bioreactor system.

### Claim 11 (depends on 10)

The bioreactor system of claim 10, wherein a cell is introduced onto the soft culture material and encapsulated in a hydrogel prior to introduction into or onto the stiff culture material.

### Claim 12 (depends on 1)

The bioreactor system of claim 1, wherein the soft culture material is introduced into or onto the stiff culture material after being loaded into the bioreactor system.

### Claim 13

A method of 3D cell passaging in a bioreactor system, comprising:
providing a population of cells to be passaged; and
introducing the population of cells into a material composition comprising two or more materials, wherein the two or more materials comprises a stiff culture material and a soft culture material that is introduced thereon to the stiff culture material and to which the cells attach.

### Claim 14 (depends on 13)

The method of claim 13, wherein the cells are introduced into the soft culture material prior to introduction of the soft culture material to the stiff culture material.

### Claim 15 (depends on 13)

The method of claim 13, further comprising causing the soft culture material to disassociate from the stiff culture material.

### Claim 16 (depends on 15)

The method of claim 15, further comprising isolating the cells and soft culture material from the material composition.

### Claim 17 (depends on 15)

The method of claim 15, further comprising recovering the dissociated cells and dividing the cells into multiple populations.

### Claim 18 (depends on 17)

The method of claim 17, further comprising introducing one or more divided cells into a second bioreactor system.

### Claim 19 (depends on 18)

The method of claim 18, wherein the cells are introduced into the second bioreactor system by introducing the cells into the material composition.

### Claim 20 (depends on 18)

The method of claim 18, wherein the cells are introduced into the second bioreactor system by introducing the cells into a second material composition contained comprising two or more materials, wherein the two or more materials comprises a stiff culture material and a soft culture material that is introduced thereon the stiff culture material to which the cells attach to at least portions of a soft culture material.

### Claim 21 (depends on 19, 20)

The method of claim 19 or 20, wherein the cells are introduced into the soft culture material prior to introduction of the soft culture material to the stiff culture material.