A primary isolation method of 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 problems such as large tissue damage, slow cell growth, and inability to large-scale quantitative production, and achieve stable culture and good cell viability.
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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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