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Method of separating derma epidermis stem cell

A technology for separation and purification of epidermal stem cells, applied in the field of expansion, separation and purification of human and mammalian skin epidermal stem cells, can solve the problem of difficult separation of epidermal stem cells and TA cells

Inactive Publication Date: 2010-05-12
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since a specific molecular marker that can accurately reflect epidermal stem cells has not been found, it is difficult to completely separate epidermal stem cells from TA cells by the above separation methods

Method used

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  • Method of separating derma epidermis stem cell
  • Method of separating derma epidermis stem cell
  • Method of separating derma epidermis stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Separation, purification and expansion of epidermal stem cells from Guanzhong dairy goats

[0033] After normal slaughter (within 30 minutes) of Guanzhong dairy goats (2.5-3.5 years old) with relatively few blood vessels in the middle of the ear, the hair was shaved and disinfected, and a piece of about 0.5×0.5cm in size was cut with ear size pliers 2 Put the skin in the normal saline containing 500 μ / ml of penicillin and streptomycin and take it back to the sterile room, rinse it several times with normal saline to remove blood stains and hair. Separate the skin from the cartilage under a dissecting microscope, and cut the skin Cut into small pieces of 0.1-0.2cm×0.1-0.2cm in size, put the epithelial side up on a glass plate with a diameter of 60mm, 3-4 pieces per plate, place on a constant temperature platform at 37°C for 15-20 minutes and then add culture Base 1-2mL, the formula of the medium is: 90% Medium199 (Gibco, LotNo.1129880) + 10% FBS (newborn bovin...

Embodiment 2

[0035] Example 2: Separation, purification and expansion of human epidermal stem cells

[0036] Take a discarded piece of about 0.5×0.5cm after the circumcision of the child in the hospital 2 Put the skin in the normal saline containing 500 U / ml of penicillin and streptomycin and bring it back to the sterile room, rinse it several times with normal saline to remove blood stains. Separate the skin from the subcutaneous tissue under a dissecting microscope, cut the skin into small pieces of 0.1-0.2cm×0.1-0.2cm in size, stick the epithelial side up on a glass plate with a diameter of 60mm, 3-4 pieces per plate, Place on a constant temperature platform at 37°C for 15 to 20 minutes, then add 1 to 2 mL of medium. The formula of the medium is: Medium19990% + 10% FBS (newborn bovine serum, Zhengzhou Baian Bioengineering Co., Ltd., Lot1120193) + 10 μg / mlinsulin (insulin, purchased from Beijing Dingguo Biotechnology Co., Ltd., http / / www.digguo.com, Sigma subpackage, original product n...

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Abstract

The invention discloses a separation and purifying method of skin epiderm stem cell, which is characterized by the following: adopting self-designed culture medium to inhibit the growth of fiber cellto growing epithelial shaped cell around the tissue; inducing surface stem cell migrate original micro-environment; gathering to grow again on the new growing epithelial shaped cell. The cloned growthcell displays obvious shaped difference from epithelial shaped cell, which is defined purifying epiderm stem cell if the detection of epiderm stem cell molecular mark is CK19, beta1-integrin,P63, alpha-6-integrin, which is positive. The invention overcomes defect of traditional enzyme digestion method, which can separate and purify human and animal epiderm stem cell.

Description

technical field [0001] The invention belongs to the technical field of separation and purification of human and mammalian adult stem cells in biomedical engineering, and specifically relates to a method for separation, purification and expansion of human and mammalian skin epidermal stem cells. Background technique [0002] Epidermal stem cells s , EpiSC s ) is a stem cell with self-proliferation and multi-directional differentiation potential. Its normal proliferation and differentiation are the basic requirements for maintaining the structural and functional integrity of the skin and its appendages (sweat glands, hair, sebaceous glands). Under physiological conditions, epidermal stem cells differentiate into a stem cell and a transit amplifying cell through asymmetric division. s TA cells), and TA cells differentiate into post-mitotic cells after multiple divisions s ) and terminally-differentiated cells s ) to supplement the need for constant renewal of epidermal cel...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08C12N5/06C12N5/071C12N5/074
Inventor 杨学义窦忠英
Owner NORTHWEST A & F UNIV
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