Method for amplifying seed cells of skin tissue engineering

Abstract

The invention relates to a method for culturing seed cells of tissue engineering. The method for amplifying the seed cells of the tissue engineering comprises the following steps of: (1) adding a macroporous microcarrier into a bioreactor, hydrating and sterilizing and then adding 50 to 100ml of fibroblast culture medium, and adding fibroblasts for culturing; (2) inhibiting the fibroblasts from amplifying when the fibroblasts are attached to the microcarrier and the growth area is near 50 percent of surface area of the microcarrier, and standing and then pouring out culture solution; and (3) washing the obtained carrier by using sterile, calcium-free and magnesium-free phosphate buffered solution (PBS), then adding an epidermal cell culture medium and epidermal cells, and continuing to stir and culture until the cells overgrow the carrier to obtain the seed cells of skin tissue engineering. The quantity and function of the cells cultured by the method can both be apparently increased,the damage of the cells is reduced, the cells can be directly used as seed cells for skin construction of the tissue engineering, and the effect is superior to those of methods using epidermal cell suspension and epidermal cell patches.

Description

technical field [0001] The invention relates to a method for culturing tissue engineering seed cells, in particular to a method for cultivating seed cells used to construct tissue engineered skin or directly applied to the treatment of burns and chronic ulcers. Background technique [0002] Tissue engineering methods have always played an important role in the treatment of burns and chronic ulcerated wounds. In the process of normal wound healing, epidermal cells play an important role. Therefore, using appropriate methods to present epidermal cells to the wound during wound healing is an important method to promote wound healing and induce local skin regeneration. Among the methods used in tissue engineering are cultured epidermal cell membranes, epidermal cell suspensions, and tissue-engineered skin that uses epidermal cells and / or fibroblasts and other scaffold materials. In 1975, Rheinwald and Green (Rheinwald JG, Green H: Serial cultivation of strains of human epiderma...

AI Technical Summary

Problems solved by technology

It needs to be combined with microcarrier culture technology when using the method of bioreactor to expand it, but because epidermal cells are terminally differentiated cells, when microcarriers are used to culture epidermal cells, due to the low adhesion ability of epidermal cells , poor anti-shear ability and rapid terminal differentiation during the culture process make the result of amplification unsatisfactory

However, the current methods of presenting keratinocytes are mostly spraying and smearing methods, without considering the combination with materials that can promote wound healing