Culture method for fibroblast derived from autologous body skin

Abstract

The invention discloses a culture method for fibroblast derived from autologous body skin. The method comprises the following steps: 1) taking autologous body skin, dividing the autologous body skin into multiple tissue blocks, attaching the tissue blocks to a culture dish, dropwise adding concentrated feta calf serum on the tissue blocks, completely soaking the tissue blocks, preventing the tissue blocks from drying and promoting the fibroblast to crawl out; 2) adding a selective culture medium into the culture dish in the step 1), placing the culture dish into an incubator to culture until the fibroblast fusion is finished, wherein the selective culture medium is used for promoting the fibroblast to grow and inhibiting other parenchyma cells to grow; 3) taking the culture dish in the step 2) out of the incubator, adding pancreatin for digestion to enable the fibroblast to fall off from the culture dish; 4) inoculating the fibroblast obtained in the step 3) into a culture bottle by a complete culture medium, enabling the fibroblast density to reach 90 percent to obtain primary fibroblast, wherein the complete culture medium is used for ending the digestion in the step 3) and is used as a culture medium for growth and proliferation of the fibroblast; 5) passaging the bottle-expanding culture of the primary fibroblast in the step 4).

Description

technical field [0001] The invention relates to the field of cell culture, in particular to a method for culturing fibroblasts derived from autologous skin. Background technique [0002] Skin fibroblasts are widely distributed in the dermis. Fibroblasts secrete a variety of substances to form the extracellular matrix, thereby forming connective tissue and exerting structural functions on the body. Fibroblasts play an important role in human life, such as guiding the formation of skin in the embryonic stage, participating in the maintenance of skin homeostasis after organ maturation, and promoting the repair of the structure and function of the damaged site. [0003] At present, the primary culture methods of autologous skin fibroblasts are roughly divided into two types: enzyme digestion method and tissue block attachment method. The enzymatic digestion method takes a long time to digest, and the operation is complicated, and because trypsin will cause irreversible damage t...

AI Technical Summary

Problems solved by technology

However, the use of enzyme digestion to remove the epidermis in the early stage will cause greater damage to fibroblasts, and the medium of this method has no selectivity for fibroblasts, and the impact of miscellaneous cells is relatively large. The purity of the cultured primary fibroblasts is low and needs to be subcultured. Fibroblasts with high purity can be obtained several times

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