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Method for inducing differentiation of sweat gland cells by epidermal stem cells and culture medium group

A technology of epidermal stem cells and induction medium, which is applied in the field of methods and medium groups, can solve the problems of epidermal stem cell differentiation and unclear metabolism mechanism, and achieve the effects of convenient and quick acquisition, good application prospects, and avoiding pollution

Active Publication Date: 2018-03-23
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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  • Method for inducing differentiation of sweat gland cells by epidermal stem cells and culture medium group
  • Method for inducing differentiation of sweat gland cells by epidermal stem cells and culture medium group
  • Method for inducing differentiation of sweat gland cells by epidermal stem cells and culture medium group

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] 1) Preparation of culture medium:

[0049] Take DMEM medium and prepare it to contain 0.5ng / mL hydrocortisone, 0.05ng / mL insulin, 1.8×10 - 4 DMEM with 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 Culture medium

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

[0051] Sweat 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 culture of epidermal stem cells

[0053] The specific operation for isolating epidermal stem cells from isolated skin tissues is as follows: take the foreskin discarded after foreskin cutting of 3-6 year old hea...

Embodiment 2

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

[0059] 1) Preparation of culture medium:

[0060] Epidermal stem cell culture medium: Take DMEM medium and prepare it to contain 0.1ng / mL hydrocortisone, 0.01ng / mL insulin, 1×10 -4 DMEM containing 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 Culture medium

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

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

Embodiment 3

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

[0065] Epidermal stem cell culture medium: Take DMEM medium and prepare it to contain 2ng / mL hydrocortisone, 1ng / mL insulin, 5×10 -4 DMEM culture with 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 to contain 100ng / mL human epidermal growth factor, 2×10 -10 mol / L cholera toxin, 5×10 -7 mol / L triiodothyronine, 8×10 -5 mol / L acetylcholine chloride in DMEM medium;

[0067] Sweat 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 differentiation of sweat gland cells from epidermal stem cells and a culture medium group thereof. Background technique [0002] The skin is the largest organ of the human body. It has the functions of protecting the body, wicking perspiration, feeling cold, heat and pressure. The sweat glands secrete sweat and emit body heat. In large-area burn patients, sweat glands are missing after the wound is repaired, which affects the body temperature regulation function. Artificial skin repairs the wound surface, with only the epidermis and dermis structure, without sweat glands and other skin appendages. It is difficult to construct sweat glands in tissue engineered skin, which is a 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 way to reconstruct...

Claims

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Application Information

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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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