摘要
目的对分别采用密度梯度离心法和贴壁筛选法分离、培养的大鼠骨髓基质干细胞(MSCs)的形态学和增殖动力学进行初步研究,以逐步完善MSCs体外培养体系。方法取1月龄SD 大白鼠胫骨和股骨骨髓,分别采用密度梯度离心法和贴壁筛选法进行体外分离、培养。倒置显微镜下进行MSCs的形态学观察和记录,MTT法测定细胞增殖度,绘制各自培养体系的传一代细胞生长曲线井比较。结果培养获得的MSCs呈梭形或多角形,表面有不规则突起。采用密度梯度离心法培养获得的细胞纯度高,生长速率较贴壁筛选法获得的细胞快,增殖显著。另外,用小牛血清培养的MSCs生长曲线与文献报道的用胎牛血清培养的MSCs生长曲线一致。结论密度梯度离心法可认为是目前体外培养MSCs的理想方法。用于体外培养MSCs的血清可能有更多选择。
Objective To make some preparation for the development of tile system of isolating and culturing bone marrow messenchymal stem cells （MSCs） in vitro, by studing the histomorphology and uhrastructure of MSCs under Gradient Centrifugation by Density and Adherence Filtration. Methods Bone marrows were collected from femur and tibia of 1 month SD rats and seeded in cell culture flask. The morphology under each isolating and culturing system were analyzed by phase contrast microscope. Cellular proliferative degree was checked with MTT method. ResultsThe doubling time of cells gained from gradient Centrifugation by Density was shorter than that of those from Adherence Filtration. But two groups had no difference on histom6rphology, both of which had spindle shape, with irregular processes on cell surface. Conclusion Gradient Centrifugation by Density should be regarded as the perfect method for isolating and culturing MSCs in vitro. It is possible to have more sorts of serums than new bovine serum in culturing MSCs.
出处
《贵州医药》
CAS
2005年第10期893-895,F0003,共4页
Guizhou Medical Journal
