间充质干细胞在构建组织工程软骨中的实验研究

Experimental Study of Mesenchymal Stem Cells in Constructing Tissue Engineering Cartilage

目的探索利用几丁质支架和间充质干细胞(MSC)构建组织工程软骨。方法抽取兔股骨骨髓,密度梯度离心分离骨髓MSC,用转化生长因子β(1TGF-β1)诱导第3代的骨髓MSC向软骨方向分化,2周后用免疫组化检测Ⅱ型胶原表达情况。将分化后的软骨样细胞传代,接种在几丁质支架上,用含TGF-β1的培养基继续培养2周,再用免疫组化检测Ⅱ型胶原表达情况,并在电镜下观察软骨细胞的生长情况。结果骨髓MSC经TGF-β1诱导后,具有软骨细胞特点,接种在几丁质支架上继续培养2周,这些细胞在支架上生长良好,并仍能很好表达Ⅱ型胶原。结论MSC经TGF-β1诱导,分化成的软骨样细胞在几丁质支架上表型稳定,生长良好。

Objective To study the methodology of constructing tissue engineering cartilage with Mesenchymal Stem Cell （MSC） and chitosan scaffolds. Methods Bone marrow was obtained from the rabbit femora, from which MSCs were separated by the method of density gradient centrifugation. TGF-β1 was used to induce chondrogenic differentiation of the third generation of MSCs. 2 weeks later,Collagen Ⅱ of the MSCs was detected by immunohistochernistry,and then the cells induced by TGF-β1 were cultured by means of chitosan scaffolds for two weeks. The expression of Collagen Ⅱ was detected again by immunohistochemistry and the cells were observed through the electronic microscope. Results After induced and cultured with TGF-β1, MSCs possessed the traits of cartilage cells. When these cartilage-like cells continued to be cultured with TGF-β1 in the chitosan scaffolds for two weeks,they grew well and expressed Collagen Ⅱ greatly. Conclusion MSCs induced into cartilage-hke cells can grow well in the chitosan scaffolds with stable chondro-phenotypes.