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Separation and culturing method of human epidermis stem cell

A technology for epidermal stem cells and epidermal cells, which is applied in the field of separation and in vitro culture of human epidermal stem cells, and can solve the problems that the characteristics of epidermal stem cells cannot be guaranteed.

Inactive Publication Date: 2006-04-26
陕西艾尔肤组织工程有限公司
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Problems solved by technology

Therefore, if the trophoblast is not used, the stem cell characteristics of epidermal stem cells during long-term culture may not be guaranteed
So far, long-term culture of human epidermal stem cells in vitro has rarely been reported at home and abroad

Method used

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  • Separation and culturing method of human epidermis stem cell
  • Separation and culturing method of human epidermis stem cell
  • Separation and culturing method of human epidermis stem cell

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Embodiment Construction

[0017] The foreskin skin of (human) healthy male after circumcision was taken to prepare epidermal cells and fibroblasts.

[0018] 1. Separation of epidermal cells (keratinocytes): take a skin 1-2 cm long and about 2 mm wide, wash it 3 times with PBS containing antibiotics; put it in protease solution, digest it at 4°C for 16 hours; take out the skin, peel off the epidermis and Dermis layer: collect epidermis slices, place in 0.25% trypsin / 0.02% EDTA (1:1) mixture, digest at 37°C for 10-15 minutes, stop digestion, pipette slightly and filter, collect cell suspension, centrifuge and discard Supernatant, add fresh culture medium, resuspend cells, adjust cell concentration to 1×10 3 / ml.

[0019] 2. Preparation of trophoblast cells: collect the above-mentioned dermis slices after peeling off the epidermis and dermis, place them in 625U / ml collagenase solution, collect the fibroblast suspension after digestion, and culture the fibroblasts until they spread about 80%. Add mitomyc...

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Abstract

The invention belongs to the technical field of biological cells, and relates to a method for separating and culturing human epidermal stem cells. The characteristics of the method include: preparation of a plate covered with extracellular matrix; separation of epidermal cells; preparation of trophoblast cells; screening and culturing of epidermal stem cells. At present, this method has been stably cultured and passaged in vitro for more than 120 PD. It has been confirmed by in vivo and in vitro experiments that the cells have no tumorigenicity, have strong proliferation ability, and can form mature full-thickness differentiated epidermis. The clinical application plays an immeasurable role, and at the same time breeds huge social and economic benefits.

Description

technical field [0001] The invention belongs to the technical field of biological cells, and relates to a method for separating and in vitro culturing human epidermal stem cells. Background technique [0002] Stem cells are a kind of primitive cells with self-renewal and differentiation potential, which can be divided into embryonic stem cells and adult stem cells. Stem cells have the ability of self-renewal and unlimited proliferation, and can produce highly differentiated functional cells. Due to the characteristics of stem cells, it provides a new way of thinking for human beings in the field of preparing autologous cells, tissues, organs, and treating difficult and miscellaneous diseases such as cancer and innate immune diseases. At present, the research on stem cells in the world mainly focuses on: the establishment of embryonic stem cells and the mechanism of induction into mature tissue cells; the induction of lateral differentiation of adult stem cells; the establis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08
Inventor 金岩董蕊
Owner 陕西艾尔肤组织工程有限公司
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