Preparation method of skin fibroblasts

Abstract

The invention discloses a preparation method of skin fibroblasts, and belongs to the technical field of biological cells. The preparation method of the skin fibroblasts comprises the following steps of: S01, specimen treatment: treating a skin specimen, and removing adipose tissues and connective tissues; S02, separation: treating the treated specimen with neutral protease, and removing epidermis to obtain dermis tissues; S03, digestion: digesting the dermis tissues by using pancreatin to obtain tissue fluid; S04, centrifugation: centrifuging the tissue fluid, and removing supernatant to obtain a cell suspension; S05, culture: transferring the cell suspension into a culture dish, adding a culture solution, and culturing; and S06, passage: culturing until the fusion degree is 75-85%, and performing passage culture. The preparation method disclosed by the invention has the advantages of simple steps, low possibility of pollution, a high cell survival rate, a good purification effect and small cell loss.

Description

technical field [0001] The invention relates to the technical field of biological cells, in particular to a preparation method of skin fibroblasts. Background technique [0002] Fibroblasts are the main components of dermal tissue and play an important role in the construction of artificial skin and tissue wound repair; the study on the culture of fibroblasts provides a powerful tool for studying the mechanism of wound healing at the cellular level. [0003] At present, for the culture and preparation of skin fibroblasts, the method adopted is as follows: the obtained specimen tissue is digested, and after the dermal cells are separated, they are centrifugally screened and cultured. [0004] However, when the epidermis and dermis are separated, the epidermal tissue is relatively dense and difficult to digest; after a long period of digestion, the cells are severely damaged, resulting in difficulty in growth and high mortality. Although it is said that reducing the digestion...

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Problems solved by technology

[0004] However, when the epidermis and dermis are separated, the epidermal tissue is relatively dense and difficult to digest; after a long period of digestion, the cells are severely damaged, resulting in difficulty in growth and high mortality

Although it is said that reducing the digestion time and increasing the frequency of cell collection can reduce the death rate of cells; however, there is still a large number of cell death, and the operation is cumbersome

Moreover, due to the poor cell activity, the cell adhesion rate is low and the growth rate is slow

[0005] At the same time, more epidermal cells are mixed in the cultured cells, and the purity of the final product is affected; additional steps are required for processing

Further lead to increased cost and increased process complexity

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