Differentiation Of Bone Marrow Mesenchymal Stem Cells Into Tracheal Cartilage Cells

Objective: To probe the optimal condition of detachment, extraction and amplification in vitro of the bone marrow mesenchymal stem cells (BMSCs) and to induce the BMSCs differentiate into chondrocytes in vitro, then identify the differentiated cells and to provide the experimental evidence for cartilagines tracheales tissue engineering .Methods: Use the bone marrow adherent cultivation to isolate and culture BMSCs. Then the cultured MSCs were induced by TGF-β1, Dexamethasone (Dex), Insulin and Vitamin C (Vit C). The induced cells were analyzed by histological observation, staining with hematoxylin-eosin staining and toluidine blue. Detect the induction of collagen type II cells (Col II) expression by RT-PCR.Results: The primary cultured MSCs adhered to plastic surface in 48 hours. After 7 days, the amount of adherent cells increased obviously and the shape of them became shuttle-shaped or polygon. After 9 days, there were small colony of cells grow. After 7 days, some of the BMSCs became irregular or ellipse from spindle shape with the function of TGF-β1, Dexamethasone (Dex), Insulin and Vitamin C (Vit C), then aggregate to form chondrogenic nodules. The induced cells exhibited the chondrocyte-like morphology by HE staining. The results of toluidine blue staining showed that the extracellular matrix was metachromatic and blue. RT-PCR indicated that Col II was expressed in the induced cells. Conclusions: The higher purity of BMSCs can be obtained by the bone marrow adherent cultivation. The BMSCs can be induced to differentiate into chondrocytes-like cells by TGF-β1, Dexamethasone (Dex), Insulin and Vitamin C (Vit C).