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Primary isolation method for hair follicle stem cells

A hair follicle stem cell and separation method technology, applied in the field of primary separation of human hair follicle stem cells, can solve the problems of large tissue damage, slow cell growth, and difficulty in adhering to the wall, and achieve stable culture and good cell viability.

Active Publication Date: 2018-12-28
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the tissue block method cannot completely remove the hair follicles from the tissue, while the collagenase or trypsin used in the enzymatic digestion method will greatly damage the tissue and affect the cultivation of stem cells
Moreover, the samples of the above-mentioned hair follicle stem cells are all from animals, and the activity of animal cells is different from that of cells in the human body. Using the above method to isolate human hair follicle stem cells, the cells grow slowly, are not easy to adhere to the wall, and cannot be mass-produced on a large scale. Some reagents are not suitable for clinical research

Method used

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  • Primary isolation method for hair follicle stem cells
  • Primary isolation method for hair follicle stem cells
  • Primary isolation method for hair follicle stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, primary separation of hair follicle stem cells

[0029] 1. Take the full-thickness scalp (30mm×5mm), rinse it several times with normal saline containing 1× double-antibody, then rinse once with 75% alcohol, and finally store it in glucose solution containing double-antibody and bring it back to the laboratory Operate within 24 hours.

[0030] 2. Cut off the hair outside the scalp and the fat and connective tissue in the subcutaneous tissue, then cut into 3-5mm wide leather strips, wash with sodium chloride injection, then wash in 75% ethanol, and finally use chlorine Sodium chloride injection was washed again.

[0031] 3. Cut the strips into tissue pieces and place them in 0.5 U / ml dispase for overnight digestion at 4°C.

[0032] 4. After 12 hours, take out the tissue block and wash it with sodium chloride injection.

[0033] 5. Take a T25 culture flask, add 1ml of 100μg / ml L-polylysine to cover the bottom of the entire culture flask, put it into a 37°...

Embodiment 2

[0036] Embodiment 2, primary separation of hair follicle stem cells

[0037] 1. Take the full-thickness scalp (30mm×5mm), rinse it several times with normal saline containing 1× double-antibody, then rinse once with 75% alcohol, and finally store it in glucose solution containing double-antibody and bring it back to the laboratory Operate within 24 hours.

[0038] 2. Cut off the hair outside the scalp and the fat and connective tissue in the subcutaneous tissue, then cut into 3-5mm wide leather strips, wash with sodium chloride injection, then wash in 75% ethanol, and finally use chlorine Sodium chloride injection was washed again.

[0039] 3. Cut the strips into tissue pieces and place them in 0.2 U / ml dispase for overnight digestion at 4°C.

[0040] 4. After 12 hours, take out the tissue block and wash it with sodium chloride injection.

[0041] 5. Take a T25 culture flask, add 1ml of 50μg / ml L-polylysine to cover the bottom of the entire culture flask, put it in a 4°C ca...

Embodiment 3

[0044] Embodiment 3, primary separation of hair follicle stem cells

[0045] 1. Take the full-thickness scalp (30mm×5mm), rinse it several times with normal saline containing 1× double-antibody, then rinse once with 75% alcohol, and finally store it in glucose solution containing double-antibody and bring it back to the laboratory Operate within 24 hours.

[0046] 2. Cut off the hair outside the scalp and the fat and connective tissue in the subcutaneous tissue, then cut into 3-5mm wide leather strips, wash with sodium chloride injection, then wash in 75% ethanol, and finally use chlorine Sodium chloride injection was washed again.

[0047] 3. Cut the strips into tissue pieces and place them in 0.3 U / ml dispase for overnight digestion at 4°C.

[0048] 4. After 12 hours, take out the tissue block and wash it with sodium chloride injection.

[0049] 5. Take a T25 culture flask, add 1ml of 70μg / ml L-polylysine to cover the bottom of the entire culture flask, put it into a 37°C...

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Abstract

The invention belongs to the field of stem cells, and discloses a primary isolation method for hair follicle stem cells. According to the method, scalp tissues are pretreated and neutral protease is added to digest the scalp tissues; after cleaning, complete hair follicles are pulled out in the hair follicle growth direction and inoculated into a culture bottle coated by L-polylysine; culture solution for stem cells are added to carry out culture; passage is carried out after the cell convergence degree is up to 80-90%. Experiments prove that the primary isolation method is capable of isolating a plenty of hair follicle stem cells, is good in cell activity and is capable of carrying out stable subsequent culture.

Description

technical field [0001] The invention belongs to the field of stem cells, and in particular relates to a primary isolation method of hair follicle stem cells, in particular to a primary isolation method of human hair follicle stem cells. Background technique [0002] As the largest organ of the human body, the skin has multiple functions such as sensation, temperature regulation, secretion and excretion, and prevention of water evaporation. The most important function is to serve as a barrier between the human body and the external environment to maintain the stability of the internal environment. important parts of. Currently, the standard treatment for skin defects is autologous skin transplantation. Due to its characteristics of no immune rejection and high survival rate, it is widely used clinically and has achieved good curative effect. However, the disadvantages of autologous skin transplantation are also very obvious. Because it is an autologous material, it itself is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0628C12N2509/00
Inventor 陈海佳葛啸虎王一飞张梦晨王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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