流动优化骨髓间充质干细胞的分化状况

背景：间充质干细胞在成体动物骨髓中含量极低。目的：观察成年大鼠骨髓间充质干细胞经生理范围流动切应力作用后,其体外生长规律、增殖及定向分化为成骨细胞及脂肪细胞的能力。方法：在体外取得SD大鼠骨髓原代间充质干细胞,利用平板流室系统加载1Pa切应力,对其进行培养及扩增,并分别向成骨细胞及脂肪细胞定向诱导。结果与结论：原代间充质干细胞为长梭形或多角形,集落样生长;经流动加载并传代后细胞生长速度加快,不以集落方式生长,而是均匀分布。细胞体积明显增大,多数为长梭形或多角形;生长曲线显示各代细胞有类似的生长规律;激光共聚焦显微镜显示CD44和CD90阳性,CD31及CD45阴性;向成骨细胞及脂肪细胞诱导14d后,VonKossa染色及油红O染色均为阳性,流动优化后骨髓间充质干细胞保持了体外大量增殖及多向分化潜能。

Multipotent differentiation status of rat bone marrow mesenchymal stem cells in vitro after shear stress loading

BACKGROUND：The number of mesenchymal stem cells in the bone marrow of adult animals is little.OBJECTIVE：To investigate whether shear stress could modify the abilities of growth,amplification and multi-potent differentiation of bone marrow mesenchymal stem cells or not.METHODS：SD rat bone marrow mesenchymal stem cells were isolated,cultured and amplified in vitro by Percoll density gradient centrifugation and adherent culture.1 Pa shear stress was loaded on primary passage mesenchymal stem cells with a flow chamber system for osteogenic and adipogenic differentiation.RESULTS AND CONCLUSION：The primary cells were mostly spindle-like in shape,like the fibroblast cells,and formatted clones.After the cells were loaded with shear stress and then passaged,they grew faster and bigger,and had the same morphologic characters similar as primary mesenchymal stem cells.The growth curves showed the growth regularity of all passage cells was similar,the laser scanning confocal microscope showed the CD41 and CD90 were positive,CD31 and CD45 were negative.After the cells were induced for 14 days,the results of Von Kossa strain and oil red O strain were both positive.The abilities of growth,amplification and multi-potent differentiation of bone marrow mesenchymal stem cells maintained after shear stress loading.