体外培养大鼠骨髓基质细胞分化为成骨细胞新方法的建立和鉴定

THE DEVELOPMENT AND IDENTIFICATION OF OSTEOBLASTS FROM RAT BONE MARROW STROMAL CELLS CULTURED in vitro

目的 改进成骨细胞骨髓培养法并进行鉴定。 方法 建立分离和培养大鼠骨髓基质细胞 (MSC)的新方法 ,并用活细胞形态学观察、HE染色、组织化学和放射免疫测定等技术进行观察和鉴定。 结果 MSC呈典型的成纤维样细胞 ,细胞生长旺盛 ,增殖能力强。体外培养第 40d后碱性磷酸酶、骨钙素量达到最高峰 ,维持至第6 0d左右开始下降。 结论 改进后的成骨细胞骨髓培养法具有分离时间缩短、污染机会减少、工作效率提高等优点 ,并且有助于MSC分化和增殖为具有良好功能特性的成骨细胞。

Objective The purpose of this study was to develop a new cultural method, so that the rat bone marrow stromal cells (MSCs) can well differentiate into osteoblasts in vitro and the cultured osteoblasts can be identified. Methods MSCs of rats were isolated and cultured, then the cultured cells were identified with inverted microscopy and undergone an HE stain for observing the morphology of living cells, and the cells were histochemically stained for examining the activity of cellular alkaline phosphatase (ALP), as well as were radio-immunologically analyzed for detecting the secretion of osteocalcin. Results The MSCs were typical fibroblast-like and possess a strang proliferative capability after cultured in vitro.The cellular ALP activity and osteocalcin secretion achieved the peak value from the fortieth day and maintained the status until the sixtieth day after cultured in vitro.Conclusion The data demonstrated that the new developed culture method has the advantages of short time, less risk of contamination and higher efficiency, and conducive to MSC's differentiating and proliferating into osteoblasts that do function well.