In-vitro culture method of chondrocytes

Abstract

The invention discloses an in-vitro culture method of chondrocytes, which comprises the following steps: cutting off cartilage tissue pieces from the surface of a joint, and repeatedly cleaning in PBS(Phosphate Buffer Solution) added with double antibodies; putting the cleaned cartilage tissue into a culture medium, cutting the cartilage tissue into pieces, and fully washing the cut cartilage tissue with PBS containing double antibodies; collecting chopped cartilage tissues, centrifuging the tissues, adding a digestive solution, and digesting the tissues; centrifugally collecting cells, adding PBS to clean the cells, adding a culture solution into a cell cluster to prepare a cell suspension, counting the cells, and adjusting the cell density; inoculating the cell, adding a low-sugar DMEMculture medium of FBS, and culturing the cells in a saturated humidity incubator; and when the cells are fused and 80-90% of the bottom of the bottle is covered, carrying out subculture. According tothe invention, the defects and defects of low cell viability, small separation quantity and poor storability in the prior art are solved to the greatest extent, and the mammary epithelial cells cultured by the method have higher cell viability and more quantity and are easier to adhere to the wall compared with the cells obtained by the traditional laboratory method.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for culturing chondrocytes in vitro. Background technique [0002] Osteoarthritis is a chronic degenerative bone and joint disease, mostly in the elderly, most of which are mainly knee joint degeneration, and the death of chondrocytes is the key to cartilage degeneration. Articular cartilage is a white, high-density connective tissue that connects joints, acts as a load-bearing material for joints, and has good friction, lubrication, and wear characteristics. Due to the lack of lymph, blood vessels, and nerves around the articular cartilage, the surrounding chondrocytes cannot proliferate rapidly, and the cartilage cannot repair itself after injury. , Widely present in joint connective tissue, but chondrocytes can rapidly proliferate and secrete extracellular matrix in a relatively suitable environment in vitro, providing raw materials for repairing damaged cartilage. [...

AI Technical Summary

Problems solved by technology

[0003] Since chondrocytes and cartilage are terminal undifferentiated cells with extremely weak proliferation ability and lack regenerative ability, self-repair is very difficult

Traditional autologous cartilage transplantation and allogeneic cartilage transplantation can achieve a certain therapeutic effect, but due to the limited source and quantity of cartilage, it is difficult to repair damaged cartilage. At present, the use of in vitro chondrocyte culture to repair damaged cartilage is becoming more and more popular.

[0004] As the only cell component in cartilage tissue, chondrocytes are an important source of cartilage tissue transplantation engineering and the key to in vitro chondrocyte transplantation. Therefore, the selection and cultivation of chondrocytes is very important, but the in vitro culture of chondrocytes is not yet mature.