体外诱导条件下大鼠骨髓基质细胞骨钙素和Ⅰ型胶原mRNA的表达

Osteocalcin and Type Ⅰ Collagen mRNA Expression of Rat Marrow Stromal Cells under Induction Condition

目的 研究体外培养大鼠骨髓基质细胞诱导条件下 ,成骨分化标志骨钙素和 型胶原 m RNA水平的表达。方法 采用成骨诱导剂 (含地塞米松 10 - 8mol/ L、β甘油磷酸钠 10 mm ol/ L和抗坏血酸 AA 5 0μg/ml)对培养状态下的不同代大鼠骨髓基质细胞进行成骨诱导 ,提取 RNA,采用 RT- PCR方法 ,以β-肌动蛋白 c DNA为内参照 ,检测骨钙素和 型胶原 m RNA的表达。结果 与对照组相比 ,接受成骨诱导剂作用的大鼠骨髓基质细胞随诱导时间的延长骨钙素和 型胶原 m RNA出现高表达 ,且维生素 D为诱导骨钙素 m RNA表达的必需成分。结论 体外培养的大鼠骨髓基质细胞保持成骨未分化状态 ,无骨钙素和 型胶原 m RNA的表达 ,诱导后可向成骨分化 ,骨钙素和 型胶原 m

Objective To investigate osteocalcin and type I collagen mRNA expression of rat marrow stromal cells under induction condition. Methods Cultured rat marrow stromal cells were induced by osteogenic compound(composed of dexamethasone 10 -8mol/L, β glyceralphosphate 10 mmol/L, ascorbic acid 50μg/ml). Osteocalcin and type Ⅰ collagen mRNA expression were measured using RT-PCR technique. Results It was found that rat marrow stromal cells in induced group have an increased time-dependent expression of osteocalcin and type Ⅰ collagen mRNA, in comparison with control groups. And the necessity of 1,25-(OH) 2D 3 in induced osteocalcin mRNA expression was confirmed. Conclusion The cultured rat marrow stromal cells kept the undifferentiated state and had osteogenic potential testified by increased osteocalcin and type I collagen mRNA expression when induced.