In-vitro cartilage cell culturing method

Abstract

The invention provides an in-vitro cartilage cell culturing method. The in-vitro cartilage cell culturing method comprises the following steps: acquiring a cartilage cell sample; performing primary culture on the cartilage cell sample; performing subculture on cartilage cells obtained after the primary culture, and when the convergence degree of the cartilage cells reaches 70-80% during the subculture, replacing a fresh culture medium and adding sulforaphane (SFN) to treat the cartilage cells. By the in-vitro cartilage cell culturing method, through the primary culture, morphs and specific molecular expression levels of the cartilage cells in the early stage are kept to be the same as those in a body to the maximum extent, so that morphologic characteristics of the cartilage cells in tight joint with a parent tissue can be displayed better; through the subculture, the cartilage cells are massively propagated, and at an appropriate time, the cartilage cells are degraded and inhibited by using the SFN, so that a degradation phenomenon during in-vitro monolayer culture can be effectively relieved, and a novel excellent method is provided for culturing the clinical cartilage cells.

Description

technical field [0001] The invention belongs to the technical field of directional culture of in vitro tissue cells, and in particular relates to a method for culturing in vitro chondrocytes. Background technique [0002] In the body, chondrocytes are the only cell type in articular cartilage, and their main function is to maintain the function of articular cartilage by secreting type II collagen and proteoglycan (aggrecan, Acan). Under normal physiological conditions, the synthesis and catabolism of chondrocytes maintain a dynamic balance; when cartilage defects and osteoarthritis (OA) occur, articular cartilage tissue is affected by various factors, and the stability between articular cartilage damage and repair The state of the cartilage is destroyed, and the catabolism of cartilage is obviously greater than the anabolism, showing that the degradation of type II collagen and proteoglycan is greater than the synthesis, in which type II collagen is usually degraded by matri...

AI Technical Summary

Problems solved by technology

The difficulty is that in the process of in vitro culture of chondrocytes, due to the loss of the specific proliferation environment in the body and the totipotency of the cells themselves, the proliferation of chondrocytes in vitro is slower and accompanied by degeneration, such as type II Decreased expression of collagen COLII, increased expression of type I collagen COLI, increased expression of MMPs, and increased expression of COLX, etc., make the chondrocytes cultured in vitro often form fibrocartilage after replantation, reduce the mechanical properties of cartilage, and lead to the effect of repairing hyaline cartilage. less than expected, becoming a major problem affecting the long-term efficacy of treatment

Images