# US 11,111,480 B2 · Culture media for multipotent stem cells

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- Number: US 11,111,480 B2
- Document ID: US11111480B2
- Status: Granted
- Kind: B2
- Application number: 16/173,962
- Filed: 2018-10-29
- Published: 2019-03-07
- Granted: 2021-09-07
- Earliest claimed: 2016-04-29
- Assignees: Hope Biosctences, LLC
- Applicants: Hope Biosciences, LLC
- Inventors: Hyeonggeun Park
- Web page: https://patentgrep.com/patent/US11111480B2

## CPC

- C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)}
- C12N 5/0665: {Blood-borne mesenchymal stem cells, e.g. from umbilical cord blood}
- C12N 5/0667: {Adipose-derived stem cells [ADSC]; Adipose stromal stem cells}
- C12N 5/0668: {Mesenchymal stem cells from other natural sources}
- C12N 2500/25: Insulin-transferrin; Insulin-transferrin-selenium
- C12N 2500/32: Amino acids
- C12N 2500/99: Serum-free medium
- C12N 2501/11: Epidermal growth factor [EGF]
- C12N 2501/113: Acidic fibroblast growth factor (aFGF, FGF-1)
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/125: Stem cell factor [SCF], c-kit ligand [KL]
- C12N 2501/31: Pituitary sex hormones, e.g. follicle-stimulating hormone [FSH], luteinising hormone [LH]; Chorionic gonadotropins
- C12N 2501/33: Insulin
- C12N 2501/39: Steroid hormones
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A growth medium for culturing mesenchymal stem cells. The medium comprises a serum, a fibroblast growth factor, and either an L-cysteine, a glutathione, or an N-acetyl cysteine. Optionally, the medium can comprise various quantities of an epidermal growth factor, a hydrocortisone, a calcium chloride, an insulin, a pituitary extract, a selenium, a stromal-derived factor, a sodium pyruvate, a transferrin, and serum-free medium.

## Claims

### Claim 1

A growth medium for culturing mesenchymal stem cells comprising:
a) a serum from about 1 percent to about 10 percent by volume;
b) a fibroblast growth factor from about 1 g/mL to about 10 ng/mL; and
c) either:
i) an L-cysteine from about 0.5 mM to about 5 mM; or
ii) a glutathione from about 0.5 mM to about 5 mM;
d) an epidermal growth factor from about 1 ng/ml to about 10 ng/ml;
e) hydrocortisone from about 10 ng/mL to about 100 ng/mL;
f) calcium chloride from about 0.01 mM to about 0.1 mM;
g) insulin from about 0.5 mg/mL to about 5 mg/mL;
h) pituitary extract from about 10 μg/mL to about 100 μg/mL;
i) selenium from about 0.1 μg/mL to about 1 μg/mL;
j) stromal-derived factor from about 1 ng/mL to about 10 ng/mL;
k) sodium pyruvate from about 2 mg/mL to about 20 mg/mL;
l) transferrin from about 0.1 mg/mL to about 1 mg/mL; and
m) serum free medium balanced to 100 percent volume; and
wherein, a cultured mesenchymal stem cell is morphologically identical to a mesenchymal stem cell of a previous generation.

### Claim 2 (depends on 1)

The stem cell growth medium of claim 1, wherein the serum is human serum.

### Claim 3 (depends on 1)

The stem cell growth medium of claim 1, wherein the serum is fetal bovine serum.

### Claim 4 (depends on 1)

The stem cell growth medium of claim 1, wherein at least one of the fibroblast growth factor or the epidermal growth factor is recombinant.

### Claim 5 (depends on 1)

The stem cell growth medium of claim 1, wherein the stromal-derived factor is recombinant stromal-derived factor.

### Claim 6 (depends on 1)

The stem cell growth medium of claim 1, wherein at least one of the insulin or the transferrin is recombinant.

### Claim 7 (depends on 1)

The stem cell growth medium of claim 1, wherein said medium contains L-cysteine.

### Claim 8 (depends on 1)

The stem cell growth medium of claim 1, wherein said medium contains glutathione.