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In-vitro inducing differentiation of umbilical cord mesenchymal stem cells into tissue engineering skin seed cells

A technology for inducing differentiation and stem cells, applied in animal cells, vertebrate cells, bone/connective tissue cells, etc., can solve the problems of unstable curative effect, cell culture and single cell therapy

Inactive Publication Date: 2010-10-20
FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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AI Technical Summary

Problems solved by technology

[0003] Previously published patents on the isolation and culture of umbilical cord mesenchymal stem cells and cell therapy are limited to cell culture and single cell therapy, and the curative effect is unstable

Method used

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  • In-vitro inducing differentiation of umbilical cord mesenchymal stem cells into tissue engineering skin seed cells
  • In-vitro inducing differentiation of umbilical cord mesenchymal stem cells into tissue engineering skin seed cells
  • In-vitro inducing differentiation of umbilical cord mesenchymal stem cells into tissue engineering skin seed cells

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Experimental program
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Embodiment Construction

[0020] With the informed consent of pregnant women who underwent caesarean section, the umbilical cord was aseptically collected, and the primary umbilical cord MSCs were cultured by enzyme digestion or tissue block attachment method. Dermal induction conditions of 5% FBS, 15 ng / mL transforming growth factor β1 (TGF-β1, R&D), 20 ng / mL bFGF, 1% ITS, 0.1 μmol / L dexamethasone, 100 U / mL penicillin and 100 μg / mL streptomycin culture medium) to induce differentiation, observe the morphological changes of the cells under an inverted phase-contrast microscope, and use the Tagman probe method to perform Real-time PCR on the induced and control cells to detect the changes in the mRNA levels of collagen type Ⅰ and type Ⅲ in the induced differentiation cells. Under specific induction conditions, the umbilical cord MSCs were induced to differentiate into epidermal stem cells. The third generation umbilical cord MSCs were digested and collected, and divided into two groups, the induction gr...

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Abstract

The invention relates to in-vitro inducing differentiation of umbilical cord mesenchymal stem cells into skin tissue engineering seed cells. The invention discloses a new research direction for human umbilical cord mesenchymal stem cells, i.e. in-vitro inducting differentiation into skin tissue engineering seed cells. The invention also discloses an experimental method thereof, which comprises the following steps: collecting the caesarean umbilical cords in aseptic environment; culturing original-generation umbilical cord mesenchymal stem cells (UCMSCs) by an enzyme digestion or tissue-piece adherent method; after purifying and subculturing to the third generation, inducing by using different in-vitro condition induction liquids to differentiate into epidermis stem cells and fibroblasts; and inducing and differentiating epidermis stem cells and fibroblasts into sudoriferous gland cells by using coculture mode with the normal sudoriferous cells. After the induced cells are digested, liquid nitrogen is stored for later use. The invention provides seed cells for skin tissue engineering, and has the advantages of strong multiplication capacity, easy inducing differentiation, low immunogenicity, abundant sources and easy obtainment.

Description

technical field [0001] The present invention relates to a new application of umbilical cord mesenchymal stem cells, in particular to research on seed cells for skin tissue engineering. The invention also relates to an experimental method for inducing differentiation of umbilical cord mesenchymal stem cells into skin seed cells in vitro. Background technique [0002] In terms of seed cells, autologous or allogeneic fibroblasts and keratinocytes are traditionally used as seed cell components to construct tissue-engineered skin. Although it has been used clinically, it has limitations such as low cell proliferation and immune rejection. Research results have shown that Apligraf, a representative product of dermal-epidermal substitutes, can only be used as a temporary cover for wounds, and cannot be used as a permanent skin substitute. Therefore, finding ideal seed cells for skin tissue engineering is the key and focus of research. Bone marrow mesenchymal stem cells (mesenchym...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 柴家科韩焱福李东杰孙天骏陶然朱桂英杨红明宋慧锋梁黎明
Owner FIRST HOSPITAL AFFILIATED TO GENERAL HOSPITAL OF PLA
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