摘要
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 inducing human bone ma rrow mesenchymal stem cells (MSCs) into chondrocytes in monolayer culture in vit ro. Methods The human bone marrow was obtained from human iliac, and the hMSCs w ere isolated by density gradient centrifuge from the mononuclear cells, and then induced into chondrogenic differentiation by adding transforming growth factor-a3 (TGF-a3), dexamethasone, ascorbic acid and so on. Results After induceme nt by special medium, the fibroblast-like hMSCs turned to be triangular or poly gonal in shape. The protein and mRNA expressions of collagen types Ⅰ, Ⅱ and Ⅲ could be detected by immunohistochemical staining and in situ hybridization respectively. The mRNAs expression of collagen types Ⅰ, Ⅱ, Ⅲ, Ⅹ, and Ⅺ and aggrecan could also be detected by RT-PCR. Conclusion The human mesenchymal st em cells can be induced to express chondrocyte matrixes, and can become the pote ntial seeding cells of articular cartilage in tissue engineering.
出处
《中华创伤骨科杂志》
CAS
CSCD
2004年第2期194-198,共5页
Chinese Journal of Orthopaedic Trauma
基金
国家高技术发展(863)计划项目(2001AA216021
2002AA205011)
国家重点基础研究发展(973)计划项目(G1999054309)
