A method for inducing and differentiating sweat gland cells from epidermal stem cells and its medium group

A technology for epidermal stem cells and induction medium, applied in the field of methods and medium groups, can solve the problems of unclear differentiation and metabolism of epidermal stem cells, and achieve the effects of convenient and fast acquisition, good application prospects and avoiding pollution.

Active Publication Date: 2020-09-25
GUANGZHOU RAINHOME PHARM&TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using epidermal stem cells to induce differentiation of sweat gland cells to achieve the purpose of expanding and culturing sweat gland cells has a good application prospect, but the cell differentiation and metabolism mechanism of epidermal stem cells is still unclear, and directed differentiation needs to overcome many uncertain factors

Method used

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  • A method for inducing and differentiating sweat gland cells from epidermal stem cells and its medium group
  • A method for inducing and differentiating sweat gland cells from epidermal stem cells and its medium group
  • A method for inducing and differentiating sweat gland cells from epidermal stem cells and its medium group

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Experimental program
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Embodiment 1

[0048] 1) Preparation of culture medium:

[0049] Take DMEM culture medium, prepare and obtain containing 0.5ng / mL hydrocortisone, 0.05ng / mL insulin, 1.8×10 - 4 mol / L adenine, 100IU / mL penicillin, 15ng / mL human epidermal growth factor, 10μg / mL transferrin, 5μg / mL glutamic acid, 5μM Y-27632 and 0.1mg / mL carboxymethyl chitosan in DMEM Culture medium;

[0050] Sweat gland cell induction medium: take DMEM medium and prepare it containing 50ng / mL human epidermal growth factor, 1×10 -10 mol / L cholera toxin, 1×10 -7 mol / L triiodothyronine, 5×10 -5 DMEM medium of mol / L acetylcholine chloride;

[0051] Sweat gland cell culture medium: take DMEM medium and prepare DMEM medium containing 50ng / mL EGF, 25mg / mL bovine pituitary extract, 100U / mL penicillin and 100μg / mL streptomycin.

[0052] 2) Acquisition and cultivation of epidermal stem cells

[0053] The specific operation of isolating epidermal stem cells from isolated skin tissue is as follows: take the foreskin discarded after ...

Embodiment 2

[0058] The difference between embodiment 2 and embodiment 1 is that

[0059] 1) Preparation of culture medium:

[0060] Epidermal stem cell culture medium: Take DMEM medium and prepare it containing 0.1ng / mL hydrocortisone, 0.01ng / mL insulin, 1×10 -4 mol / L adenine, 50IU / mL penicillin, 5ng / mL human epidermal growth factor, 2μg / mL transferrin, 1μg / mL glutamic acid, 1μM Y-27632 and 0.01mg / mL carboxymethyl chitosan in DMEM Culture medium;

[0061] Sweat gland cell induction medium: take DMEM medium and prepare it containing 25ng / mL human epidermal growth factor, 0.1×10 -10 mol / L cholera toxin, 0.5×10 -7 mol / L triiodothyronine, 2×10 -5 DMEM medium of mol / L acetylcholine chloride;

[0062] Sweat gland cell culture medium: take DMEM medium and prepare DMEM medium containing 10 ng / mL EGF, 10 mg / mL bovine pituitary extract, 50 U / mL penicillin and 50 μg / mL streptomycin.

Embodiment 3

[0064] The difference between embodiment 3 and embodiment 1 is that 1) the preparation of medium:

[0065] Epidermal stem cell culture medium: take DMEM medium and prepare it containing 2ng / mL hydrocortisone, 1ng / mL insulin, 5×10 -4 DMEM culture of mol / L adenine, 200IU / mL penicillin, 50ng / mL human epidermal growth factor, 50μg / mL transferrin, 10μg / mL glutamic acid, 20μM Y-27632 and 1mg / mL carboxymethyl chitosan base;

[0066] Sweat gland cell induction medium: take DMEM medium and prepare it containing 100ng / mL human epidermal growth factor, 2×10 -10 mol / L cholera toxin, 5×10 -7 mol / L triiodothyronine, 8×10 -5 DMEM medium of mol / L acetylcholine chloride;

[0067] Sweat gland cell culture medium: take DMEM medium and prepare DMEM medium containing 100ng / mL EGF, 50mg / mL bovine pituitary extract, 200U / mL penicillin and 200μg / mL streptomycin.

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Abstract

The invention discloses a method for inducing differentiation of sweat gland cells by epidermal stem cells, wherein the method comprises the following steps: 1) separating epidermal stem cells from isolated skin tissues, and subculturing with an epidermal stem cell culture medium; 2) subculturing the epidermal stem cells obtained in the step 1), adding a sweat gland cell induced culture medium, and carrying out induced differentiation; and 3) subculturing and proliferating by the sweat gland cell culture medium. The invention also provides a corresponding culture medium group. In the method, the cell source effectively avoids the ethical controversy. The preparation method is helpful to preparation of the sweat gland cells with homogeneous cell directional differentiation and proliferationconditions.

Description

technical field [0001] The invention relates to the technical field of biological tissue culture, in particular to a method for inducing and differentiating sweat gland cells from epidermal stem cells and a culture medium group thereof. Background technique [0002] The skin is the largest organ in the human body and has functions such as protecting the body, perspiration, and sensing heat, cold, and pressure. Sweat glands secrete sweat to dissipate body heat. For patients with large area burns, after the wound is repaired, the sweat glands are missing, which affects the thermoregulation function. The artificial skin repairs the wound, only has the structure of the epidermis and dermis, without sweat glands and other skin appendages. It is difficult to construct sweat glands in tissue-engineered skin, which is a difficult problem to be solved at present. Simulating the mechanism of sweat glands to induce stem cells to differentiate into sweat gland cells may be the only w...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0633C12N2500/25C12N2500/32C12N2500/34C12N2500/40C12N2500/84C12N2501/01C12N2501/11C12N2501/39C12N2501/395C12N2501/727C12N2501/805C12N2506/09
Inventor 黄燕飞车七石刘少辉
Owner GUANGZHOU RAINHOME PHARM&TECH CO LTD
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