骨髓间充质干细胞的分离培养及生物学特性研究

目的：通过体外细胞的培养方法将骨髓间充质干细胞由骨髓血中分离出来并加以纯化，进一步在体外培养条件下研究其增殖及生长特征，从而为探讨后续利用组织工程学方法修复肌肉骨骼系统组织缺损的可能性提供实验基础。方法：从骨髓血中提取间充质干细胞，体外培养扩增，再以相差显微镜、电镜观察并绘制生长曲线。结果：（1）通过离体培养，可以使体内环境下低丰度的骨髓间充质干细胞实现数目扩增。（2）体外培养条件下骨髓间充质干细胞具有成纤维细胞的生长特性。（3）体外单层培养条件下贴壁生长的骨髓间充质干细胞会出现“返祖现象”。（4）骨髓间充质干细胞内细胞器与其生物活性变化相一致。结论：骨髓间充质干细胞体外培养成功为今后利用自体间充质干细胞通过组织工程学方法修复肌肉骨骼系统的组织缺损提供功能细胞奠定了基础。

THE ISOLATION,CULTURE OF THE MSCs OF BONE MARROW AND STUDY ON THEIR BIOLOGICAL FEATURES IN VITRO

ObjectiveTo investigate the behavior of MSCs in vitro.Methods:MSCs were extracted from bone marrow and purified.The isolated cells were observed with phase-contrast microscope and electronic microscope.Mesenchymal stem cells were extracted from the bone marrow in blood and were proliferated in vitro.Then they were observed under the phase-contrast microscope and electronic microscope and the growth curve was drawn accordingly.Results:(1)MSCs that are in low abundance can grow quickly when cultured in vitro.(2)MSCs have the tendency to be the fibroblasts.(3)When cultured in mono-layer,those attached MSCs showed actavism after some subculture.(4)The structures of the organellae varied with the changes of cell biological characteristics.Conclusion:The success of in vitro culture of MSCs will be useful for amendment of skeletal tissue defects by tissue engineering aftermath.