CULTURE MEDIUM OF ADIPOSE-DERIVED STEM CELL, METHOD FOR PREPARING THE SAME, AND COMPOSITION INCLUDING THE SAME FOR PROMOTING HAIR GROWTH

• US 20150037435A1
• Filed: 08/29/2012
• Published: 02/05/2015
• Est. Priority Date: 03/09/2012
• Status: Active Grant

• Current Assignee: Changwon National University Industry Academy Cooperation Corps
• Original Assignee: Changwon National University Industry Academy Cooperation Corps
• Inventors: Lee, Gwang, Kim, Dong Wan, Seo, Mi Jung, Joo, Woo Hong, Kim, Sun Hee
• Granted Patent: US 9,999,589 B2
• Time in Patent Office: Days
• US Class Current: 424/574
• CPC Class Codes: A61K 2800/70 Biological properties of the composition as a whole
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 8/981 of mammals or bird
• CPC Class Codes: A61Q 7/00 Preparations for affecting hair growth
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2500/84 from mammals
• CPC Class Codes: C12N 2506/1384 from adipose-derived stem cells [ADSC], from adipose stromal stem cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0653 Adipocytes; Adipose tissue
• CPC Class Codes: C12N 5/0667 Adipose-derived stem cells [ADSC]; Adipose stromal stem cells

Abstract

Provided are a culture medium of an adipose-derived stem cell, a method for preparing the same, and a composition for promoting hair growth, in which the composition includes the culture medium. The adipose-derived stem cell (ADSC-T) according to the present invention exhibits long lifespan, improved cell proliferation rate, and extended proliferation period, as compared with a primary adipose-derived stem cell (ADSC), and thus, the adipose-derived stem cell (ADSC-T) can be usefully used for the study about the adipose-derived stem cell and the mass production of the culture medium of the adipose-derived stem cell. In addition, according to the present invention, the culture medium of the adipose-derived stem cell (ADSC-T) that expresses a T antigen of SV40 exhibits excellent hair growth effectiveness and can be usefully used as a raw material for the hair loss prevention and hair growing agents.

Claims

1. A method for preparing a culture medium of an adipose-derived stem cell (ADSC-T), the method comprising:

(a) culturing the adipose-derived stem cell after isolating the adipose-derived stem cell through a centrifuge after treating an enzyme, collagenase, to a suctioned adipose tissue;

(b) preparing the adipose-derived stem cell (ADSC-T) expressing a T antigen of SV40 by transfecting a plasmid expression vector (pEF321β

-T) into the adipose-derived stem cell cultured in the above step (a); and

(c) obtaining a culture medium by culturing the adipose-derived stem cell (ADSC-T) prepared as described above.

2. The method according to claim 1, wherein in the above step (a), the collagenase and the suctioned adipose tissue are mixed in a weight ratio of 1:

1 to react them at 30 to 40°

C. for 40 to 50 minutes.

3. The method according to claim 1, wherein in the above step (a), the centrifugation is carried out at 500 to 1000 G for 2 to 5 minutes.

4. The method according to claim 1, wherein in the above step (a), the isolated adipose-derived stem cell is cultured in Dulbecco'"'"'s Modified Eagle Media (DMEM) added with 10% Fetal Bovine Serum (FBS), 100 units/ml of penicillin, and 100 μ

g/m of streptomycin in an incubator of 37°

C. and 5% CO₂.

5. The method according to claim 1, wherein in the above step (b), the transfection is carried out by using one or more methods selected from the group consisting of a calcium phosphate transfection, an electroporation, a microinjection, and a liposome injection.

6. The method according to claim 1, wherein in the above step (c), the culture medium is a supernatant obtained by collecting the culture medium after cell confluence reached 70 to 90% in the incubator, centrifuging the culture medium, and then removing cellular residue from the culture medium.

7. A culture medium of the adipose-derived stem cell (ADSC-T) prepared according to the method described in any one of claims 1 to 6.

8. A pharmaceutical composition for promoting hair growth, the pharmaceutical composition comprising the culture medium of the adipose-derived stem cell (ADSC-T) described in claim 7 as an effective component.

9. A cosmetic composition for promoting hair growth, the cosmetic composition comprising the culture medium of the adipose-derived stem cell (ADSC-T) described in claim 7 as an effective component.