Induction medium and induction method for stem cell differentiation into sweat gland-like cells

A technology for inducing culture medium and sweat gland-like cells, applied in the field of stem cells, can solve the problems of unclear differentiation mechanism, unclear composition of conditioned medium, uncertain sweat gland-like cells, etc., and achieves easy popularization, low cost, and culture equipment. simple effect

Active Publication Date: 2021-04-30
福州市皮肤病防治院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The sweat gland-like cells were subjected to heat shock treatment, and the sweat gland-like cells were directly co-cultured with the mesenchymal stem cells after the heat shock, or the culture supernatant of the sweat gland-like cells after the heat shock was collected and co-cultured with the mesenchymal...

Method used

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  • Induction medium and induction method for stem cell differentiation into sweat gland-like cells
  • Induction medium and induction method for stem cell differentiation into sweat gland-like cells

Examples

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Embodiment 1

[0063] Example 1 Sweat gland-like cell induction medium

[0064] This embodiment provides a sweat gland-like cell induction medium, the sweat gland-like cell induction medium is DMEM / F-12 cell culture medium and also includes the following components, and the dosage of each component is:

[0065] Fetal Bovine Serum (FBS): 5%

[0066] Epidermal growth factor: 50ng / mL

[0067] Basic fibroblast growth factor: 50ng / mL

[0068] Recombinant human ectoderm dysplasia A1 protein (Ectodysplasin A1 Protein): 20ng / mL

[0069] Recombinant human Wnt3a protein: 20ng / mL

[0070] Bone Morphogenetic Protein 4: 20ng / mL

[0071] Hepatocyte Growth Factor: 25ng / mL

[0072] Triiodothyronine: 2ng / mL

[0073] Hemisuccinyl hydrocortisone: 0.15 μg / mL

[0074] Insulin-Transferrin-Selenite: 1%

[0075] L-Glutamine: 1 μmol / mL

[0076] Penicillin and streptomycin (1:1): 100U / mL

Embodiment 2

[0077] Example 2 Application of sweat gland-like cell induction medium in inducing stem cells to differentiate into sweat gland-like cells

[0078] The stem cells used in this example are human umbilical cord mesenchymal stem cells (hUC-MSCs), and the steps for obtaining hUC-MSCs include:

[0079] A. Operating table After the human umbilical cord is obtained, it is submerged in 0.9% sterilized saline, and aseptically transported to the laboratory.

[0080] B. Take it out in the ultra-clean bench, wash 3 times with PBS buffer containing 1% double antibody (penicillin and streptomycin), to remove the blood ulcer and fully clean the lumen of the umbilical vein, strip off the arteriovenous, take the umbilical cord connective tissue, cut 4-5mm 3 The size of the organization block.

[0081] C. Inoculate the tissue block in a 6-well cell culture plate, add 0.5-l mL of DMEM / F-12 medium containing 10% fetal bovine serum to the culture plate, and place the culture plate in a cell cult...

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Abstract

The invention discloses an induction medium for differentiating stem cells into sweat gland-like cells, and belongs to the technical field of stem cells. A induction culture medium is a DMEM/F-12 cell culture medium containing fetal bovine serum, an epidermal growth factor, a basic fibroblast growth factor, a recombinant human ectoderm dysplasia A1 protein, a recombinant human Wnt3a protein, a bone morphogenetic protein 4, a hepatocyte growth factor, triiodothyronine and hemisuccinyl hydrocortisone. The composition is prepared from insulin- transferrin-sodium selenite, L-glutamine and penicillin/streptomycin double antibiotics. The sweat gland induction culture medium can effectively induce stem cells, especially mesenchymal stem cells, to be differentiated into sweat gland-like cells with sweat gland-like cell phenotypes in vitro, the sweat gland-like cells can form a tubular structure similar to a sweat gland structure in glue, and the requirement for sweat glands in the treatment process of large-area burn patients can be met.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to an induction medium and an induction method for stem cells to differentiate into sweat gland-like cells. 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. Among them, sweat glands, as important functional skin appendages, can regulate body temperature, secrete sweat, and excrete some metabolites of the human body. Through the secretion and evaporation of sweat, sweat glands can discharge 25% of the body's heat to maintain a relatively constant body temperature. The number of sweat glands in the human body is certain, and its development is a complicated process. Once a large area is damaged, it will not be possible to quickly and effectively restore functional sweat glands. [0003] In mild burns, sweat gland-like cells can rely on the de...

Claims

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Application Information

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IPC IPC(8): C12N5/071
CPCC12N5/0633C12N2506/1346C12N2506/1392C12N2501/11C12N2501/115C12N2501/415C12N2501/155C12N2501/12C12N2500/25C12N2501/30C12N2500/32Y02A50/30
Inventor 吴小末梅沁
Owner 福州市皮肤病防治院
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