中国组织工程研究与临床康复2008，Vol.12Issue(38)：7577-7581,5.
不同分离方法对骨髓间充质干细胞成软骨分化的影响
Effects of different isolation methods on chondrogenic differentiation of bone marrow mesenchymal stem cells
摘要
Abstract
BACKGROUND: At present, there has been no definite experiment systemically evaluating adherent separation and density gradient centrifugation to isolate bone marrow mesenchymal stem cells (BMSCs). Whether the two methods produce different influences on BMSC induction and differentiation remains unclear.OBJECTIVE: This study was designed to verify difference of these two isolation methods in chondrogenic differentiation of BMSCs.DESIGN, TIME AND SETTING: A controlled observation was performed at the Shengjing Hospital Affiliated to China Medical University from March to September 2005. MATERIALS: Twenty Japanese big-ear rabbits, aged 2-3 months, weighing 1.2-2.0 kg, were included for this study. METHODS: BMSCs were isolated by adherent separation and density gradient centrifugation. Two groups of BMSCs were taken from the same passage and induced towards chondrogenic differentiation with transforming growth factor beta 1. MAIN OUTCOME MEASURES: Growth of BMSCs was observed under an inverted microscope to draw growth curves; Type II collagen expression was detected by immunohistochemistry. Type II collagen mRNA expression was determined by in situ hybridization. RESULTS: The growth curves demonstrated that cellular growth velocity of the two groups tended to be the same. Immunohistochemistry results showed that the efficiency of adherent separation and density gradient centrifugation for promoting chondrogenic differentiation of BMSCs was 76.1% and 77.7%, respectively, and in situ hybridization results showed that the efficiency was 70.3% and 71.0%, respectively. No significant difference in differentiation efficiency existed between the adherent separation and the density gradient centrifugation. CONCLUSION: Adherent separation and density gradient centrifugation had no different influences on BMSC growth and chondrogenic differentiation.关键词
骨髓/间充质干细胞/Ⅱ型胶原/软骨分化/分离培养方法/贴壁分离法/密度梯度离心法分类
医药卫生引用本文复制引用
王广斌,付勤,杨礼庆,付永慧..不同分离方法对骨髓间充质干细胞成软骨分化的影响[J].中国组织工程研究与临床康复,2008,12(38):7577-7581,5.基金项目
辽宁省教育厅项目(05L568)the Grant from Project of Department of Education of Liaoning Province, No. 05L568 （05L568）
