大鼠骨髓和脂肪来源间充质干细胞的成骨特性比较

Osteogenic property of rat bone marrow mesenchymal stem cells versus adipose mesenchymal stem cells

背景:尽管骨髓来源和脂肪来源的间充质干细胞均具有向骨和软骨分化潜能,但终究带有各自来源组织的特点,那么是否意味着两者在非基因影响下成骨特性存在一定差异呢?目的:比较大鼠骨髓和脂肪来源的间充质干细胞在体外成骨分化中的特性区别。设计、时间及地点:细胞学体外对照观察,于2007-07/2008-03在中国科学院昆明动物研究所国家级重点实验室完成。材料:6周龄和18周龄SD大鼠各2只,分别用于制备骨髓间充质干细胞和脂肪间充质干细胞。方法:选取第3代骨髓间充质干细胞和脂肪间充质干细胞,按5×107L-1接种后各分为2组:诱导组加入含10-8mol/L地塞米松、10mol/Lβ-甘油磷酸、50mg/L维生素C的成骨诱导培养基1.5mL,未诱导组不加入成骨诱导培养基。主要观察指标:碱性磷酸酶染色与茜素红染色结果,碱性磷酸酶和骨钙素定量分析结果。结果:成骨诱导7,14d,经诱导的骨髓基质干细胞和脂肪间充质干细胞呈碱性磷酸酶阳性,有钙结节形成,未诱导组呈阴性。成骨诱导3,7,10d,未诱导组间比较碱性磷酸酶及骨钙素含量均基本相似(P>0.05);与未诱导组比较,脂肪间充质干细胞诱导组碱性磷酸酶及骨钙素含量均明显升高(P<0.05),骨髓间充质干细胞诱导组升高幅度更为显著(P<0.001)。结论:大鼠骨髓来源间充质干细胞体外成骨性能强于脂肪来源间充质干细胞,在骨组织工程种子细胞的选用中应优先考虑前者。

BACKGROUND： Both adipose mesenchymal stem cells （AMSCs） and bone marrow mesenchymal stem cells （BMSCs） can differentiate into osteoblasts and chondrocytes, but different stem cells from diverse tissues take characteristics of the source organizations. In a non-genomic pattern, both have some differences in osteogenic properties. OBJECTIVE： To compare differences in osteogenic differentiation between AMSCs and BMSCs in vitro. DESIGN, TIME AND SETTING： The cytology in vitro controlled experiment was performed at the State Key Laboratory, Institute of Kunming Animal, Chinese Academy of Sciences from July 2007 to March 2008. MATERIALS： Two 6-week-old and two 18-week-old Sprague Dawley rats were selected to prepare BMSCs and AMSCs, respectively. METHODS： The third passage of BMSCs and AMSCs at a density of 5×10^7 L^-1 were divided into two groups. Cells in the induction group were incubated in 1.5 mL of osteoinductive medium, supplemented with 10^-8 mol/L dexamethasone, 10 mol/L β-glycerophosphoric acid, and 50 mg/L vitamin C. Cells in the non-induction group were not treated with osteoinductive medium. MAIN OUTCOME MEASURES： Results of alkaline phosphatase staining and alizarin red staining; Results from quantitative analysis of alkaline phosphatase and osteocalcin expression. RESULTS： At 7 and 14 days following osteogenic induction, a significant positive reaction to alkaline phosphatase staining was detected, with calcium nodus. A negative reaction was detected in the non-induction group. At 3, 7 and 10 days, alkaline phosphatase and osteocalcin expression was similar in the non-induction group （P 〉 0.05）. Compared to the non-induction group, alkaline phosphatase and osteocalcin expression was significantly increased in the AMSCs induction group （P 〈 0.05）. There were significant increase in the BMSCs induction group （P 〈 0.001）. CONCLUSION： Osteogenic property of BMSCs was higher than that of AMSCs. BMSCs should be selected in preference as seed cells in bone tissue engineering.