大鼠骨髓基质干细胞的成骨诱导及鉴定

Osteogenic inducement and identification of mesenchymal stem cells in rats

目的:研究大鼠骨髓基质干细胞(BMSCs)在体外培养、诱导后的形态学改变及生物活性,探讨BMSCs作为骨组织工程种子细胞的可能性和可行性。方法:通过改良的骨髓培养法分离成年大鼠BMSCs,经条件培养液诱导培养后,进行形态学观察和碱性磷酸酶(ALP)、骨钙素(OCN)的测定。结果:经过体外成骨诱导的BMSCs表现出增殖、聚集、结节和矿化期的阶段性形态特征,同时在诱导2周后表达ALP活性,在结节期和矿化期OCN表达呈阳性。结论:体外成骨定向诱导的BMSCs具有典型的成骨细胞的形态和功能性特征,可以作为骨组织工程的种子细胞。

Objective：To observe the morphological changes and phenotype characteristics of bone marrow stromal stem cells （BMSCs） during osteogenic inducement and to identify the feasibility as the seed cells for tissue engineering of bone. nethod：BMSCs were isolated by modified method from adult rats, Osteogenic medium was used to stimulate BMSCs into osteoblast-like cells. The phenotype was examined with morphological method,and the levels of alkaline phosphatase （ALP） and osteocalcin （OCN） were detected.Result：The osteogenic stimulated BMSCs represented the morphological change of osteoblast,phases of proliferation,aggregation,nodulation and mineralization could be observed. These cells also represented osteoblast-like cells functionally by an increased expression of ALP after two weeks of osteogenic stimulation,secretion of OCN during the phase of nodulation and formation of mineralized cellular nodes.Conclusion：BMSCs after osteogenic stimulation show the phenotype characteristics of osteoblast,and can be used for the seed cells of tissue engineering of bone.