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目的研究人骨髓间充质干细胞(humanmarrowmesenchymalstemcells,hMSCs)在体外单层培养条件下诱导分化为软骨细胞的可行性。方法取志愿者骨髓9例,密度梯度离心获得hMSCs,进行诱导培养。光镜下观察诱导细胞形态学的改变,免疫组化、原位杂交、RT-PCR等方法检测胶原和糖蛋白的体外表达情况。结果经诱导后hMSCs形态由长梭形逐渐向多角形转变,原位杂交、免疫组化等检测到Ⅰ、Ⅱ、Ⅲ型胶原表达,RT-PCR检测到Ⅰ、Ⅱ、Ⅲ、Ⅹ、Ⅺ型胶原、aggrecan等mRNA表达。结论hMSCs单层诱导培养条件下,能分泌软骨细胞特征性细胞外基质如Ⅱ型胶原、aggrecan等,具有作为软骨组织工程种子细胞来源的可能。
Objective To study the feasibility of human mesenchymal stem cells (hMSCs) differentiating into chondrocytes under monolayer culture in vitro. Methods 9 cases of bone marrow from volunteers, density gradient centrifugation to obtain hMSCs, induction culture. The morphological changes of induced cells were observed under light microscope. The expression of collagen and glycoprotein in vitro was detected by immunohistochemistry, in situ hybridization and RT-PCR. Results The morphological changes of hMSCs from long fusiform to polygonal, in situ hybridization and immunohistochemistry showed that the expression of type Ⅰ, Ⅱ and Ⅲ collagen was detected by RT-PCR. The expression of type Ⅰ, Ⅱ, Ⅲ, Ⅹ and Ⅺ Collagen, aggrecan and other mRNA expression. Conclusion Under the condition of monolayer induction, hMSCs can secrete chondrocytes characteristic extracellular matrix such as type Ⅱ collagen, aggrecan and so on, which may be used as the source of cartilage tissue engineering seed cells.