摘要
目的 摸索体外分离培养骨髓间充质干细胞的最佳条件 ,并观察其部分生物学活性。方法 采用常规的细胞培养传代技术以及光、电镜技术和细胞增殖活性检测法 ,观察不同贴壁时间、不同浓度血清、不同种植密度对骨髓间充质干细胞生长、增殖、形态的影响 ,并观察培养细胞的部分生物学特性。结果 以选用 4～ 2 4h贴壁的有核细胞 ,加入 5 %～ 1 0 %胎牛血清、种植密度(4～ 8)× 1 0 4个 /ml的培养条件为最适宜细胞生长 ;在此条件下 ,培养扩增的细胞仍具有干细胞多向分化性 ;其形态呈梭状 ,具有快速增殖的能力 ;培养细胞在 3～ 4d进入对数生长期 ,随后进入平台期 ,分裂相细胞明显减少 ;光镜下细胞呈梭状或类成纤维状 ,2～1 0代的细胞呈均质状 ;超微结构显示为早期幼稚细胞形态。结论 建立了骨髓间充质干细胞体外分离、培养的条件 ,探讨了骨髓间充质干细胞部分生物学特性 。
Objective To observe some biological features of bone marrow mesenchymal stem cells and explore the best conditions for isolating and culturing in vitro . Methods Common cell culture technique, light and electron microscopy were used to study the effects of the growth, proliferation, morphology of the bone marrow mesenchymal stem cells in different adherent time, concentration of serum and cell density. Results The best culture condition in vitro for growth was 4-24 hours adherent time, 5%-10% fetal bovine serum, (4-8)×10 4/ml cell density. The cells were spindle in shape and had a strong ability of proliferation. The time for cell duplication was 3 to 4 days. The cells showed the characteristics of stem cells in electron microscope. Conclusion The best condition for isolation and culture of bone marrow mesemchymal stem cells was successfully established and some biological features were obserred. It found a base for further investigation and using of mesenchymal stem cells.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2001年第5期553-555,共3页
Journal of Third Military Medical University
基金
国家重点基础研究发展规划项目! ("973"项目 ) (G1 9990 542 0 5)
