# US 10,370,637 B2 · Technology for sustaining pluripotency and improved growth of stem cells in culture

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- Number: US 10,370,637 B2
- Document ID: US10370637B2
- Status: Granted
- Kind: B2
- Application number: 15/380,811
- Filed: 2016-12-15
- Published: 2017-06-15
- Granted: 2019-08-06
- Earliest claimed: 2015-12-15
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: Prudence Talbot; Sabrina Lin; Antonio Loza
- Web page: https://patentgrep.com/patent/US10370637B2

## CPC

- C12N 5/0606: {Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696)}
- C12N 2501/999: Small molecules not provided for elsewhere
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A method of culturing pluripotent stem cells is provided. The method includes culturing pluripotent stem cells in a pluripotent stem cell culture medium supplemented with an additive, where the additive includes a source of acetate ions, a carboxylic acid, or a physiologically acceptable salt of the carboxylic acid, or a combination of these substances, in an amount effective to maintain the pluripotent stem cells in culture in an undifferentiated pluripotent state. Also included are pluripotent stem cell culture media and methods of making such media.

## Claims

### Claim 1

A method for improving the culturing of pluripotent stem cells in a pluripotent stem cell (PSC) culture medium, comprising supplementing the PSC culture medium with 5 mM to 10 mM sodium acetate and culturing the pluripotent stem cells in the supplemented PSC culture medium; wherein the additive is effective to maintain pluripotency of the pluripotent stem cells to a greater extent as compared to control pluripotent cells cultured in the PSC culture medium not supplemented with the additive.

### Claim 2 (depends on 1)

The method of claim 1, wherein the pluripotency of the pluripotent stem cells is maintained in culture for 20 or more passages.

### Claim 3 (depends on 1)

The method of claim 1, wherein the cultured pluripotent stem cells attach to and spread faster on culture substrate, do not differentiate with repeated passaging, contain fewer dying cells, are more motile as cells or colonies, or grow faster, or any combination thereof, as compared to the control pluripotent cells.

### Claim 4

A method of rescuing a culture of pluripotent stems cells that has begun differentiating, comprising supplementing a PSC culture medium with 10 mM sodium acetate and incubating the culture of pluripotent stems cells in the supplemented culture medium; wherein the additive is effective to reduce or prevent further differentiation of the culture of pluripotent stems cells as compared to control pluripotent cells cultured in the PSC culture medium not supplemented with the additive.

### Claim 5

A method of creating induced pluripotent stems cells, comprising supplementing a PSC culture medium with 5 mM to 10 mM sodium acetate and culturing reprogramming cells in the supplemented PSC culture medium; wherein the additive is effective to increase production of induced pluripotent stem cells derived from the reprogramming cells as compared to control reprogramming cells cultured in the PSC culture medium not supplemented with the additive, the source of acetate ions comprising acetic acid, a physiologically acceptable salt of acetic acid, or a metabolic precursor of acetic acid, or a combination thereof.

### Claim 6

A method of improving a PSC culture medium, comprising supplementing the culture medium with 5 mM to 10 mM sodium acetate wherein the additive is effective to maintain pluripotency of pluripotent stem cells, reduce or prevent further differentiation of a culture of pluripotent stem cells that has begun differentiating, or increase induction of pluripotent stem cells from reprogramming cells, as compared to respective control cells cultured in the PSC culture medium not supplemented with the additive.