Method for screening and culturing extracellular hair follicle stem cell matrix for clinic treatment level cell therapy

Abstract

The invention discloses culturing, screening and amplifying techniques for hair follicle stem cells for clinic treatment level cell therapy. The bottlenecks are always research hotspots and difficulties in the biological stem cell field; currently, by carrying out separation and purification by virtue of remarkable adhering properties of a basement membrane, the hair follicle stem cells are obtained by virtue of three-dimensional culture and amplification measures. By depending on amplification anchorage-dependent cells, a three-dimensional high-simulation in-vivo extracellular matrix system is provided for screening and separating the levels of target cells. The method is a novel culture technique for culture in vitro of biological cells of stem cells of adults (embryo skin) in a high-simulation in-vivo adhesion environment. Skin seed hair follicle cells of targeted tissue engineering cultured through adhesion screening in the environment of a high-simulation substrate have high abdication capacity and multiplication capacity and can form a differentiated cortex structure in vitro. By screening immunogenic cells, immune escape is safely achieved, the transplant time is prolonged, the clinic application of tissue engineering is increased, the healing of wounds is promoted, and a solution with absolute advantages for the clinic treatment level cell therapy is provided for diseases of skin.

Description

1. Technical field: [0001] The invention belongs to the field of biological stem cell technology and tissue engineering. It relates to a technique for constructing a highly simulated in vivo attachment environment in vitro, separating and screening, and cultivating and expanding hair follicle stem cells. Obtain active seed stem cells from hair follicle stem cells used in tissue engineering skin construction technology. 2. Background technology: [0002] The skin is the largest organ of the human body. It plays a very important physiological role in resisting biological invasion, ultraviolet radiation, preventing water loss, regulating body temperature and maintaining individual appearance. It is an important part of the human immune system, and skin cells are the composition The basic unit of this organ, the basic research field of skin cells is an important scientific research way to reveal skin health, development, prevention and treatment. [0003] The extracellular mat...

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Problems solved by technology

Therefore, if the trophoblast is not used, it is possible that the differentiation of skin cells during long-term culture may cause uncontrollable development of subsequent research;

[0036] Method 6 (homologous fibroblast attachment screening method): Scholar Jin Yan et al. used references to improve scholars HagarB and Dunnwald mouse fibroblasts, changed the donor fibroblasts pre-expanded in vitro as the attachment substrate as a trophoblast, and cultured homologous fibroblasts. Epidermal cells, but the patent does not specify the detailed steps of how to prepare the trophoblast plate, the plating cycle is short, the fibroblast adhesion force is low, and the repeated PBS cleaning and purification process of screening will lose part of the cultured fibroblasts successfully attached to the target, thus Reduce the yield of target skin cells. Skin cells are theoretically homogeneous. Another technical problem is that the complexity of the culture medium should be considered when two kinds of cells share the same culture environment. The two kinds of cells may have competitive growth and proliferation. Inhibition and other phenomena, the subsequent isolation of epidermal stem cells is technically difficult, and it is not superior to the existing six common methods. It is suggested that this method can be changed into a compound recipe and added to other screening materials with good effect of covering the dish and greater adhesion behavior. Target stem cell yield, or inactivated fibroblastic plated cells

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