An Experimental Study On The Chondrogenic Differentiation From Rabbit Bone Marrow Mesenchymal Stem Cells Transfected By Transforming Growth Factor-β1 Gene In Three Dimensional Culture In Vitro

Objective: To discuss the effect of induceing the BMSCs to the chondrogenic differentiation by gene transfection and three dimensional culture in vitro . Methods: To investigate its biological features and proliferation with BMSCs cultured in different condition between monolayer culture and three dimensional culture in vitro. Construction of cosmid vector inserting TGF-β1 gene (pAdEasy1.TGF-β1) and replication-deficient recombinant adenovirus coding for TGF-β1 gene (Ad. TGF-β1). To study the chondrogenic differentiation from bone marrow mesenchymal stem cells transfected by Ad.TGF-β1 gene in three dimensional culture in vitro. Results: We could get the bone marrow mesenchymal stem cells (BMSCs) isolated from bone marrow by separating and fulling bone marrow and bone cavity from long bone of four weeks newland rabbits.The BMSCs were purified through a method of the density gradient ultracentrifuge, and they were suscessfully cultured in monolayer and three dimensional culture system in vitro.The more steady three dimensional culture system was successfully established. This provide a probability to get a great number of seeding cells for the cartilage tissue engineering. An safe,efficient and reliable method of constructing recombinant Ad vectors was established. The BMSCs transfected with TGF-β1 could persistently secrete marked increase of specific extracellular matrix(ECM) of chondrocyte such as Collagen Ⅱ and aggrecan,which was more excelled in three dimensional culture system than that in monolayer culture. Conclusions: The BMSCs can be better induced to the chondrogenic differentiation by gene transfection and three dimensional culture system in vitro .