摘要
【目的】研究大鼠骨髓间充质干细胞(MSCs)的分离纯化和在诱导条件下的成骨能力。【方法】取SPF级SD大鼠6只,采用全骨髓贴壁培养法分离成年大鼠骨髓间充质干细胞,取传至第3代的MSCs进行细胞形态和免疫组化鉴定;应用含地塞米松、β-甘油磷酸钠和维生素C的诱导分化培养液诱导传代细胞向成骨细胞分化,检测碱性磷酸酶(ALP)的活性和细胞矿化作用进行成骨细胞鉴定。【结果】全骨髓贴壁培养法能有效分离纯化大鼠骨髓MSCs,MSCs在含体积分数为10%胎牛血清的低糖DMEM(L-DMEM)培养液中生长性状相对稳定,增殖速度快;诱导条件下,细胞ALP活性明显增高,并出现了矿化结节。【结论】建立了一种体外分离纯化、培养扩增大鼠骨髓MSCs的方法,成骨能力肯定,为中药蛋白质组学研究提供材料基础。
Objective To investigate the isolation and purification of rat bone marrow mesenchymal stem cells（MSCs） and to explore the osteoblastic capacity after induction.Methods MSCs were obtained from 6 specific pathogen free（SPF） rats.The bone marrow adherent method was used to isolate the adult rat MSCs.The third generation of subcultured MSCs was identified by cell morphological observation and immunohistochemical method.Then the fluid containing dexamethasone,β-sodium glyerophosphate and vitamin C was used to induce MSCs to differentiate into osteoblast.The activity of alkaline phosphatase（ALP） and the cell mineralization nodules were detected for osteoblastic identification.Results The bone marrow adherent method can isolate and purify MSCs effectively.MSCs can grow smoothly and proliferate rapidly in the L-DMEM culture with 10% fetal bovine serum added.Under induction condition,ALP activity increased obviously and mineralization nodule presented.Conclusion A method for the isolation,purification and proliferation of rat MSCs is established successfully,which shows a certain osteoblastic capacity and will supply evidence for proteomics of Chinese herbal medicine.
出处
《广州中医药大学学报》
CAS
2008年第2期176-179,共4页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家自然科学基金资助课题(编号:30273631)
