Constructing tissue-engineered cartilage with porcine bone marrow mesenchymal stem cells in vitro / 中国组织工程研究

ABSTRACT

AIM:Repair of trachea is disturbing the surgeon. Tissue engineering technology will probably resolve this problem. Seed cell is one of the key factors in engineered tracheal cartilage construction. This study investigated the feasibility of constructing tissue-engineered cartilage from porcine bone marrow mesenchymal stem cells(MSCs) cultured and induced in vitro using tissue engineering technique. METHODS:The experiment was performed in the Central Laboratory of Peking Union Medical College Hospital between October 2006 and May 2007. ①By density gradient centrifugation,the MSCs were isolated and purified from porcine bone marrow. The MSCs had been cultured and induced in the defined medium mainly including transforming growth factor-?1,and then the type Ⅱ collagens were detected by immunohistochemical assay. The induced MSCs were seeded onto polyglycolic acids(PGA) scaffold as experimental group,and PGA scaffold were implanted into subcutaneous tissue as control group. The cell-scaffold construct was wrapped around a silicon tube(0.4 cm in diameter) and implanted into subcutaneous tissue of porcine. All specimens were harvested after in vivo culture for 6,8 and 10 weeks and evaluated by gross view,histology,and immunohistochemistry. RESULTS:①The MSCs were obtained by density gradient centrifugation method,and abundant seed cells were obtained after culture and amplification. ②The MSCs differentiated towards chondrocyte when cultured in the specific medium in vitro and were verified by the positive result of collagen type Ⅱ through immunohistochemistry. ③After implanted into subcutaneous tissue for 6,8 and 10 weeks,the cell-scaffold formed a tubular cartilage,which was very similar to normal porcine tracheal cartilage in both gross view and histology. And the result of collagen type Ⅱ through immunohistochemistry was positive. CONCLUSION:The in vivo and vitro cultured MSCs from porcine bone marrow can generate tissue-engineered cartilage under chondrogenic induction.