摘要
目的 研究体外培养的猪骨髓间质干细胞 (BoneMarrowStemCells,MSCs)在特定培养液作用下向软骨细胞表型转化 ,探讨其作为组织工程化软骨的种子细胞的可行性。方法 取成年崇明长枫杂交猪髂骨骨髓 5ml,在低糖DMEM完全培养液培养 2周 ,传代后以高糖DMEM无血清特定培养液诱导 (含胰岛素 2mg L、转铁蛋白 3mg L、丙酮酸 10 0mg L、地塞米松 10 - 7mol L、TGF β110ng ml) ,在相差显微镜和电镜下进行观察 ,免疫组化检测Ⅱ型胶原分泌 ,原位杂交检测Ⅱ型胶原mRNA表达。结果 细胞形态由成纤维样梭形向多角形、多边形转变 ,透视电镜观察见大量扩张粗面内质网、高尔基体、线粒体。诱导培养后第 7,14dⅡ型胶原免疫组化阳性 ,同时原位杂交检测Ⅱ型胶原mRNA表达呈阳性。结论 MSCs在特定培养液诱导下能向软骨细胞表型转化 ,并能分泌软骨特异性基质 ,有可能成为软骨组织工程较理想的种子细胞来源的应用前景。
Objective\ To investigate the feasibility of chondrogenic phenotype differentiation of adult swine bone marrow stem cells(MSCs) in a defined medium as seeding cells in cartilage tissue engineering. Methods\ A volume of 5 ml bone marrow was aspirated from swine iliac crest and cultured in the complete medium of DMEM LG for two weeks. The growth and ultrastructure of the cultured MSCs were observed. Immunohistochemistry and in situ hybridization were applied to detect the expression of collagen type Ⅱ. Results\ The MSCs changed from a spindle like fibroblastic appearance to a polygonal shape when transferred from the complete medium of DMEM LG to a defined medium. A large amount of endoplasmic reticulum was observed in large Golgi complex and mitochondria. The differentiation of MSCs toward chondrogenic phenotype was verified by the positive result of collagen type II through immunohistochemistry and in situ hybridization respectively. Conclusions\ Bone marrow stem cells obtained from adult swine can differentiate to be chondrogenic phenotype when cultured in vitro. MSCs can likely be served as optimal autogenous cell source for cartilage tissue engineering.
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2002年第1期12-14,共3页
Chinese Journal of Plastic Surgery
基金
国家重点基础发展规划项目 (973
G19990 5 43 0 5 )资助
