# US 12,215,352 B2 · Cell culture method for mesenchymal stem cells

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- Number: US 12,215,352 B2
- Document ID: US12215352B2
- Status: Granted
- Kind: B2
- Application number: 18/124,346
- Filed: 2023-03-21
- Published: 2024-06-13
- Granted: 2025-02-04
- Earliest claimed: 2015-03-04
- Assignees: Mesoblast International Sarl
- Applicants: Mesoblast International Sarl
- Inventors: Paul Simmons; Colby Suire
- Web page: https://patentgrep.com/patent/US12215352B2

## CPC

- C12N 5/0662: {Stem cells}
- C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)}
- C12N 2500/90: Serum-free medium, which may still contain naturally-sourced components
- C12N 2500/99: Serum-free medium
- C12N 2501/11: Epidermal growth factor [EGF]
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/135: Platelet-derived growth factor [PDGF]
- C12N 2502/115: Platelets, megakaryocytes
- 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 relates to a cell culture medium that promotes cell proliferation during serum-free cell culture.

## Claims

### Claim 1

A serum-free cell culture medium comprising:
a basal medium;
10 ng/ml or less of platelet derived growth factor-BB (PDGF-BB);
1 ng/ml or less of fibroblast growth factor 2 (FGF2); and 5 ng/ml or less of epidermal growth factor (EGF); and
mesenchymal lineage precursor cells in an undifferentiated state.

### Claim 2 (depends on 1)

The culture medium of claim 1, wherein the level of FGF2 is from 2 pg/ml to 1 ng/ml.

### Claim 3 (depends on 1)

The culture medium of claim 1, wherein the level of FGF2 is 1 ng/ml.

### Claim 4 (depends on 3)

The culture medium of claim 3, wherein the level of PDGF-BB is 10 ng/ml.

### Claim 5 (depends on 4)

The culture medium of claim 4, wherein the level of EGF is 5 ng/ml.

### Claim 6 (depends on 1)

The culture medium of claim 1, wherein the level of PDGF-BB is from 3.0 ng/ml to 10 ng/ml.

### Claim 7 (depends on 1)

The culture medium of claim 1, wherein the level of EGF is from 0.08 ng/ml to 5 ng/ml.

### Claim 8 (depends on 1)

The culture medium of claim 1, wherein the level of EGF is from 3 ng/ml to 5 ng/ml.

### Claim 9 (depends on 1)

The culture medium of claim 1, wherein the basal medium is alpha-minimal essential medium.

### Claim 10 (depends on 1)

The culture medium of claim 1, wherein the culture medium is xeno-free.