OBJECTIVE: To investigate the feasibility of constructing tissue engineered cartilage by inducing bone marrow-derived mesenchymal stem cells (MSCs) in centrifuge-tube culture.METHOD: By Percoll density gradient centrifuge and monolaryer culture in vitro MSCs were isolated from rabbit bone marrow and were purified. Some MSCs were seeded into centrifuge tube culture and allowed to form three-dimensional aggregates in a chemically defined medium mainly including transforming growth factor- b1 (TGF-beta1, 10 ng/ml), dexamethasone (10-7), ascorbic acid (50 microg/ml), 1% insulin, 1% selenium and 1% transferring. At day 7, 14, 21, aggregates were take out and specific markers of differentiation of cartilage were detected by the appearance of toluiding blue merachromasia and the immunohistochemistry detection of type II collagen and in situ hybridization detection of collagen type II mRNA.RESULT: MSCs seeded into centrifuge tube culture became analogous cartilage tissue after 7-21 days, their specific markers of chondrogeneses were positive in the appearance of toluiding blue merachromasia and the immunohistochemistry detection of type II collagen and in situ hybridization detection of collagen type II mRNA.CONCLUSION: MSCs have the capacity to undergo chondrogenic differentiation.
Animals Cell Differentiation