Novel method for in-vitro separated culture of human epidermal cells

Abstract

The invention provides a novel method for the in-vitro separated culture of human epidermal cells. The method comprises the steps: taking normal human skin tissue containing epidermis and dermis, and carrying out in-vitro digestion and separation, so as to obtain the human epidermal cells; and then, carrying out in-vitro amplified culture on the epidermal cells. According to the method provided by the invention, fat is not required to be removed, and the epidermis and the dermis are not required to be separated; skin tissue taken by the method provided by the invention is all normal human skin tissue, including scalps with hair, tissue with fat and the like; the method is simple, convenient and fast, is not restricted to time and is applicable to the direct separation of different skin tissue, such as the scalps with hair and the tissue with fat; and an epidermal cell bank with relatively high activity can be established, so that the requirements on clinical transplantation are facilitated.

Description

technical field [0001] The invention relates to a new method for separating and culturing human epidermal cells in vitro, belonging to the field of histocytology. Background technique [0002] Epidermal cells are the epithelial cells on the surface of the skin, mainly composed of keratinocytes, dendritic cells and Michael cells, which play a vital role in wound healing, especially the lack of epidermal cells in severe burn patients and the duration of the disease, concurrent Therefore, the use of tissue engineering technology to separate, cultivate and preserve epidermal cells in vitro, and to form composite skin directly or with other materials for clinical application has always been a priority for burn workers. Research hotspots. [0003] The culture and reproduction of epidermal cells began with the relevant literature reports published by Rheinwald in 1975, and then explored by some scholars. At present, there are many methods such as Tr method, Tr-C method, Dispase me...

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Problems solved by technology

However, since 3T3 cells may produce and express metabolites and antigens that are unfavorable to the human body, and the fetal bovine serum albumin in the culture medium can be combined with the cultured epidermal cell membrane, it is easy to cause rejection after skin grafting, and the medium containing serum is easy to spoil , affecting the observation of cell proliferation and morphology. In addition, this method requires two kinds of culture medium at the same time, and the culture medium needs to add more than 10 kinds of additional components. Individual components such as cholera toxin, selenium ion, epidermal growth factor, etc. are expensive and difficult to buy; 3T3 The degree of cell inactivation is difficult to grasp and the operation is complicated, which is not conducive to popularization and application. At present, it is only used for primary culture or is eliminated; the air-liquid interface culture method is a relatively new method of epidermal cell culture. It is the application of allogeneic epidermis or similar In the dermis-like substance to increase the contact between the basal layer cells of the epidermis and the culture medium, and expose most of the superficial cells to the air, the cultured keratinocytes can be completely differentiated under this artificially created physiological condition

It can cultivate epidermal cells similar to normal skin structure, with a high degree of keratinization, but the operation process is relatively complicated, requiring dermal analogues such as HSE, and the cultivation process is difficult to observe, and the cultivation period is long, which is not conducive to mass cultivation

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