体外诱导小鼠骨髓基质干细胞向成骨细胞分化相关基因表达的研究  

The gene expressions involved in osteogenic differentiation of bone marrow stromal cells of mice in vitro

作　　者：王琳[1] 刘德瑜[1] 石心泉[1] 裴开颜[1] 李东[1] 贾孟春[1] 

机构地区：[1]国家人口计生委科学技术研究所,北京100081

出　　处：《中华创伤骨科杂志》2006年第11期1062-1066,共5页Chinese Journal of Orthopaedic Trauma

基　　金：国家自然科学基金(30400251)

摘　　要：目的体外培养小鼠骨髓基质干细胞(BMSCs)诱导分化为成骨细胞,定量分析其骨生成关键标志基因的表达水平。方法体外培养小鼠BMSCs 7、14、21 d,提取细胞总RNA,采用实时荧光定量RT-PCR方法检测Ⅰ型胶原、碱性磷酸酶(ALP)、骨桥蛋白(OPIN)和骨唾液酸蛋白(BSP)基因的表达水平。结果小鼠BMSCs体外稳定增殖和分化,Ⅰ型胶原培养14 d和21 d mRNA水平分别是培养7 d时的0．75±0．04倍和(0．34±0．03)倍;ALP、OPN和BSP基因表达水平随着培养时间延长而上升,其中培养21 d时的ALP、OPN和BSP mRNA水平分别是培养7 d时的(19．70±2．36)倍、(150．12±9．31)倍和(7．73±0．58)倍。结论荧光定量RT-PCR可以灵敏地检测成骨细胞关键标志基因表达水平,随着培养时间延长,Ⅰ型胶原的表达水平逐渐下降,而ALP、OPN和BSP的表达水平显著上升。Objective To qualitatively analyze the expressions of key marker genes involved in osteogenic differentiation of bone marrow stromal cells （BMSCs） of mice at defined stages in vitro using real-time RT-PCR. Methods The BMSCs of mice were cultured in vitro and induced to osteoblasts for 7, 14, and 21 days. The total RNA was extracted from the cultured cells and the gene expression levels of collagen Ⅰ , bone alkaline phosphatase （APL）, osteopontin （OPN） and bone sialoprotein （BSP） in the osteoblasts at the defined stages were determined by real-time RT-PCR. Results The Col Ⅰ mRNA levels in 14 and 21 days were （0.75 ± 0.04） and （0.34 ±0. 03） times respectively of that in 7 days. ALP, OPN and BSP mRNA levels in 21 days were （ 19.70 ± 2.36）, （ 150. 12 ± 9.31） and （7.73 ± 0. 58） times of those in 7 days respectively. Conclusions Col Ⅰ expression levels tend to gradually decline while ALP, OPN and BSP levels gradually increase along with the culture time. Real-time RT-PCR is a reliable method for investigation of gene expressions in BMSCs cultured in vitro.

关 键 词：骨髓基质干细胞 基因表达 实时荧光定量RT—PCR 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]