论文部分内容阅读
目的 建立一种体外分离、培养和鉴定小鼠骨髓间质干细胞的方法 ,探讨体外培养中间质干细胞的一些生物学特点 ,为利用MSCs促进创伤修复提供实验基础。方法 分离 5～ 6周龄的小鼠胫骨、股骨 ,用预冷的IMDM培养基冲洗出骨髓 ,经密度梯度离心得到骨髓单个核细胞 ,接种后 1 2～ 1 6d形成单层贴壁的成纤维状细胞。体外多向诱导分化鉴定分离的细胞 ,用传代的细胞进行生长曲线的测定、观察其接种贴壁率、检测细胞周期和超微结构。结果 体外传代培养的MSCs具有分化形成成骨和成脂细胞的能力 ;MSCs倍增时间约为 3 8h ,传代后 1 2h贴壁达 90 %以上 ,细胞周期显示有 80 %细胞处于G0 G1 期 ,并用电镜照片显示其超微结构。结论 在本实验条件下 ,体外培养的MSCs具有多向分化潜能 ,体外生长稳定 ,传代后的细胞适应性强 ,增殖较快 ,超微结构和细胞周期表现出较早期细胞特点。可望用于自体创伤促愈
OBJECTIVE: To establish a method for the isolation, cultivation and identification of mouse bone marrow mesenchymal stem cells in vitro and to explore some biological characteristics of the mesenchymal stem cells in vitro. It provides an experimental basis for using MSCs to promote wound healing. Methods The tibia and femur of 5 ~ 6-week-old mice were isolated and the bone marrow was washed out with pre-cooled IMDM medium. The bone marrow mononuclear cells were obtained by density gradient centrifugation and formed into monolayer adherent fibroblasts 12 ~ 16 days after inoculation Cells. Induction of differentiation in vitro identification of isolated cells, with passaged cells growth curve was measured to observe the inoculation rate, cell cycle and ultrastructure. Results MSCs differentiated into osteoblasts and adipocytes were differentiated in vitro. The doubling time of MSCs was about 38 h, and the adherent cells reached more than 90% at 12 h after passage. The cell cycle showed that 80% of cells were in G0 G1 phase Electron microscopy shows its ultrastructure. Conclusion Under the experimental conditions, MSCs cultured in vitro have multidirectional differentiation potential, stable growth in vitro, strong adaptability to cells after passage, rapid proliferation, and ultrastructural and cell cycle characteristics of early-stage cells. Expected to be used for self-healing wounds