兔骨髓间质干细胞的成骨诱导及鉴定

Osteogenic Stimulation and Identification of Mesenchymal Stem Cells in Rabbits

目的 :探讨体外成骨诱导的兔骨髓间质干细胞 ( MSCs)的形态和功能特征 ,以及其作为骨组织工程种子细胞的可行性。方法 :选用生长状态良好的兔骨髓间质干细胞 ,在体外进行成骨活性诱导 ,并进行形态学观察和碱性磷酸酶、骨钙素的测定。结果 :经过体外成骨诱导的兔 MSCs表现出增殖、聚集、结节和矿化期的阶段性形态特征 ,同时在刺激 1周后表达碱性磷酸酶活性 ,在结节期和矿化期表达骨钙素活性。结论 :体外成骨诱导的的兔 MSCs具有典型的成骨细胞的阶段性特征和功能性特征 。

Objective: To observe the phenotype characteristics of mesenchymal stem cells(MSCs) during osteogenic stimulation and to identify the feasibility as the seed cells for tissue engineering of bone. Methods: Osteogenic medium (expansion medium with 10 mmol·L -1 dexamethasone,50 μg·L -1 ascorbic acid and 10 mmol·L -1 β glycerophosphate ) was used to stimulate MSCs into osteoblast like cells. The phenotype was examined with morphological method, and the levels of alkaline phosphatase (ALP) and osteocalcin were determined. Results: The osteogenic stimulated MSCs represented the morphological developent of osteoblast, phases of proliferation, aggregation, nodules and mineralization could be observed. These cells also represented osteoblast like cells functionally by an increased expression of ALP after one week of osteogenic stimulation, secretion of osteocalcin during the phase of nodules and formation of mineralized cellular nodes. Conclusion:MSCs after osteogenic stimulation show the phenotype characteristics of osteoblast, and can be used for the seed cells of tissue engineering of bone.