# US 2014/0154800 A1 · Embryonic stem cell line and method for preparing the same

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- Number: US 2014/0154800 A1
- Document ID: US20140154800A1
- Status: Published application
- Kind: A1
- Application number: 14/091,106
- Filed: 2013-11-26
- Published: 2014-06-05
- Earliest claimed: 2003-12-30
- Assignees: H. Bion Co., Ltd.
- Applicants: H. Bion Co., Ltd.
- Inventors: Sung-Il ROH; Woo-Suk HWANG; Byeong-Chun LEE; Sung-Keun KANG; Young-June RYU; Eu-Gene LEE; Soon-Woong KIM; Dae-Kee KWON; Hee-Sun KWON; Ja-Min KOO; Eul-Soon PARK; Youn-Young HWANG; Hyun-SOO YOON; Jong-Hyuk PARK; Sun-Jong KIM
- Web page: https://patentgrep.com/patent/US20140154800A1

## CPC

- C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N 5/0606: {Pluripotent embryonic cells, e.g. embryonic stem cells [ES] (embryonic germ cells C12N5/0611, induced pluripotent stem cells C12N5/0696)}
- C12N 15/8776: {Primate embryos}
- C12N 5/0619: {Neurons}
- C12N 5/0623: {Stem cells}
- C12N 2500/25: Insulin-transferrin; Insulin-transferrin-selenium
- C12N 2500/38: Vitamins
- C12N 2501/115: Basic fibroblast growth factor (bFGF, FGF-2)
- C12N 2501/13: Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
- C12N 2501/235: Leukemia inhibitory factor [LIF]
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

An embryonic stem cell line derived from a nucleus-transferred oocyte prepared by transferring a nucleus of a human somatic cell into an enucleated human oocyte may differentiate into various desired cell types.

## Claims

### Claim at position 1

1-19. (canceled)

### Claim 20

A neuro progenitor differentiated from an embryonic stem cell line derived from a nucleus-transferred oocyte prepared by transferring a nucleus of a human somatic cell into an enucleated human oocyte.

### Claim 21 (depends on 20)

The neuro progenitor of claim 20, wherein the embryonic stem cell line is a cell line deposited under the accession number of KCLRF-BP-00092.

### Claim 22 (depends on 20)

A method for preparing the neuro progenitor of claim 20, comprising the steps of:
(1) culturing the embryonic stem cell line to form an embryoid body;
(2) culturing the embryoid body in the presence of an agent suitable for differentiating a cell of the embryoid body into the neuro progenitor; and
(3) selecting a cell expressing a marker of the neuro progenitor and culturing the selected cell to obtain the neuro progenitor.

### Claim 23 (depends on 22)

The method of claim 22, wherein the embryonic stem cell line is a cell line deposited under the accession number of KCLRF-BP-00092.

### Claim 24 (depends on 22)

The method of claim 22, wherein the agent employed in step (2) is selected from the group consisting of retinoic acid; ascorbic acid; nicotinamide; N-2 supplement; B-27 supplement; and a mixture of insulin, transferrin, sodium selenite and fibronectin.

### Claim 25 (depends on 3)

A medium for use in carrying out the in vitro culturing in step (4) of claim 3, comprising:
95 to 110 mM NaCl; 7.0 to 7.5 mM KCl; 20 to 30 mM NaHCO3; 1.0 to 1.5 mM NaH2PO4; 3 to 8 mM sodium lactate; 1.5 to 2.0 mM CaCl2.2H2O; 0.3 to 0.8 mM MgCl2. 6H2O; 0.2 to 0.4 mM sodium pyruvate; 1.2 to 1.7 mM fructose; 6 to 10 mg/ml human serum albumin; 0.7 to 0.8 μg/ml kanamycin; 1.5 to 3% essential amino acids; 0.5 to 1.5% nonessential amino acids; 0.7 to 1.2 mM L-glutamine; and 0.3 to 0.7% a mixture of insulin, transferrin and sodium selenite.

### Claim 26 (depends on 25)

The medium of claim 25, which comprises:
99.1 to 106 mM NaCl; 7.2 mM KCl; 25 mM NaHCO3; 1.2 mM NaH2PO4; 5 mM sodium lactate; 1.7 mM CaCl2.2H2O; 0.5 mM MgCl2.6H2O; 0.3 mM sodium pyruvate; 1.5 mM fructose; 8 mg/ml human serum albumin; 0.75 μg/ml kanamycin; 2% essential amino acids; 1% nonessential amino acids; 1 mM L-glutamine; and 0.5% a mixture of insulin, transferrin and sodium selenite.