Method for extracting active components of skin stem cells

A technology of skin stem cells and active ingredients, applied in the field of extraction of skin stem cell active ingredients, can solve the problems of low survival rate of skin stem cells, small number of skin stem cells, long digestion time, etc., achieve cell structure integrity, reduce digestion time, and relieve stem cell the effect of damage

Inactive Publication Date: 2017-04-26
SHANXI MEDICAL UNIV
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Problems solved by technology

The number of skin stem cells extracted by this method is small, and the digestion time of collagenase III and trypsin is long, which causes greater damage to the cells, resulting in low survival rate and poor activity of the extracted skin stem cells, which ultimately affects its efficacy in treating hair loss

Method used

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  • Method for extracting active components of skin stem cells
  • Method for extracting active components of skin stem cells
  • Method for extracting active components of skin stem cells

Examples

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Embodiment 1

[0023] Example 1: Isolation and culture of skin stem cells.

[0024]Take the human skin tissue and wash it with normal saline. After the swelling fluid and the skin tissue are naturally layered, discard the solution below; add an equal volume of normal saline to the skin tissue, place it on a shaker and shake it for 5 minutes, and let it stand. Leave for 1 to 5 minutes, discard the solution below; then add an equal volume of normal saline, centrifuge at 1000rpm for 5 minutes, and discard the supernatant; repeatedly wash the tissue with normal saline and discard the supernatant until the supernatant is clear and there is no blood in the tissue. Obtain epidermal tissue.

[0025] Take 0.25g type Ⅰ collagenase and 0.25g type Ⅱ collagenase, add to 200mL normal saline to dissolve evenly, then add 2g PMSF protease inhibitor to dissolve evenly, adjust the pH value of the solution at 7.2-7.4 to obtain mixed digestive juice.

[0026] Mechanically crush the cleaned epidermal tissue, add...

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Abstract

The invention discloses a method for extracting active components of skin stem cells. The method comprises the steps of adding human skin tissues to a normal saline mixed digestive fluid of a type I collagenase and a type II collagenase added with a PMSF protease inhibitor for constant temperature shaking table culture; stopping digestion by a culture medium M199 containing 10% by volume of fetal calf serum; performing centrifuging and precipitate collection; performing cell re-suspension of normal saline to obtain compound skin stem cells; and screening out Dlk1<+>/Scal<-> skin stem cells by applying magnetic bead marked Dlk antibody and Scal antibody. The purity of the Dlk1<+>/Scal<-> skin stem cells extracted with the method can reach 95.8%; and the skin stem cells are large in quantity, high in cell survival rate and high in activity, are applied to drugs for treating hair loss, and have a remarkable hair growth effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for extracting active ingredients of skin stem cells. The active ingredients of skin stem cells extracted by the method of the invention can be used for treating hair loss. Background technique [0002] Alopecia is a common and frequently-occurring disease in dermatology, mainly in men, which seriously affects people's appearance and psychological state. There are many types of hair loss, with androgenic sources being the most common. The prevalence of hair loss in China is about 30.29%, and 25% of men aged 25 to 55 have androgenetic alopecia. And Western men's hair loss is more obvious, and the hair loss rate of men between the ages of 30 and 70 is positively correlated with age. With the increase of social pressure, men's psychological burden increases, hair loss is developing towards a younger age, and the prevalence rate is also increasing year by year. [0003] Many s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/074A61K35/36A61P17/14
CPCC12N5/0625A61K35/36C12N2500/32C12N2500/84C12N2501/20C12N2509/00
Inventor 于保锋孟凡秀张琪解军曹睿温晓薇杨巧艳加三三温丽敏白欣艳姚志坚曾思衡袁洋洋张紫燕张丽娜王萱田九博马培元刘志贞傅松涛杨丽红郭睿王惠珍徐钧刁海鹏陈显久胡晓年弓韬王海龙赵虹
Owner SHANXI MEDICAL UNIV
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