大鼠骨髓间充质干细胞体外向成骨细胞分化培养的生物学特性  

Biological characteristics of in vitro differentiation of bone marrow stromal cells into osteoblasts in rats

作　　者：李雪峰[1] 刘亮[2] 王东[2] 

机构地区：[1]北京市丰台区长辛店医院骨科,北京市100043 [2]山西医科大学第二附属医院骨科,山西省太原市030000

出　　处：《中国组织工程研究与临床康复》2011年第23期4185-4188,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:骨髓间充质干细胞因取材方便、对机体损伤小,是目前最为合适的骨组织工程种子细胞,但其体外培养的具体方法及生物特性仍无定论。目的:建立一种简单有效的骨髓间充质干细胞的体外培养及成骨诱导方法,并探讨其体外培养的生物学特点。方法:采用贴壁法分离提纯Wistar大鼠骨髓间充质干细胞,传至第3代细胞后,分别在普通培养基(对照组)和成骨诱导培养基(实验组)中培养10d,绘制细胞生长曲线。于培养第4,7,10,13,16天检测两组细胞碱性磷酸酶活性,并对细胞爬片行Vonkossa染色。结果与结论:采用贴壁法获得了均一性较好的骨髓间充质干细胞,对照组细胞生长速度明显快于实验组细胞。对照组细胞碱性磷酸酶活性明显低于实验组(P<0.05)。实验组细胞Vonkossa染色为阳性,对照组为阴性。说明贴壁筛选法是一种简单有效的骨髓间充质干细胞的分离纯化方法,骨髓间充质干细胞在体外可诱导分化为成骨细胞。BACKGROUND：Because of convenience and small injury to the body,bone marrow stromal cells（BMSCs） are the most appropriate seed cells in bone tissue engineering. But the methods in vitro cultivation and the biological characteristic are not clear. OBJECTIVE：To establish a simple and effective BMSCs cultured in vitro and osteoblasts induction method,and to explore the biological characteristics in vitro. METHODS：The BMSCs of Wistar rats were seperated and purified by using of adherent method. The passage cells of the third generation were cultured in the common medium（control group） and osteogenic induction medium（experimental group） for 10 days,the growth cure of each group was drew. The alkaline phosphatase activities（APA） of two groups were detected at 4,7,10,13,16,days＇ culture and cover glass was treated with Von Kossa staining. RESULTS AND CONCLUSION：The better homogenicity of BMSCs were obtained by adherent method. The growth velocity of the cells cultured in the control group was obviously faster than that in the experimental group. The APA in control group was significantly lower than that in experimental group（P 0.05） . The Von Kossa＇ staining in the experimental group was positive and negative in the control group. Adherent screening method is a simple and effective method to separate and purify BMSCs. BMSCs can be differentiated into osteoblasts in vitro.

关 键 词：体外培养 骨髓间充质干细胞 生物学特性 诱导分化 干细胞培养 

分 类 号：R394.2[医药卫生—医学遗传学]