兔骨髓间充质干细胞的分离培养及其生物学性状的研究

目的：分离培养兔骨髓间充质干细胞，对其生物学性状进行观察。方法：用密度梯度离心法与贴壁培养法相结合，分离兔骨髓间充质干细胞，进行体外培养扩增，绘制其生长曲线，用倒置显微镜、细胞爬片、扫描电镜、透射电镜观察细胞的生物学性状。结果：密度梯度离心后，活细胞比例在95％以上，贴壁培养的细胞呈纺锤形，细胞增殖力强，平均倍增时问为32h，细胞的增殖能力随传代次数的增加而有所下降。结论：密度梯度离心与贴壁培养相结合可以提高细胞分离效率。体外培养的兔骨髓问充质干细胞生长稳定，增殖力强，可以作为组织工程的种子细胞。

Study of the isolation culture and biological characteristics of rabbit bone marrow-derived mesenchymal stem cells in vitro

Objective:To establish a method to isolate and culture rabbit bone marrow-derived mesenchymal stem cells(BMSCs) and investigate their biological characteristics.Methods:Bone marrow aspirate of New Zealand white rabbit was density gradient centrifuged and cultured in plastic culture bottles.The cells were examined by invert microscope,scanning and transmission electron microscope.The growth curve was drawn.Results:95% cells were alive after density gradient centrifugation.BMSCs had a similar spindle-like morphology.The average time for cell doubling was 32 hours.The proliferation ability declined with time of the culture.Conclusion:The combination of density gradient centrifugation and adhere culture is an effective method to isolate BMSCs from bone marrow aspirate.BMSCs cultured in vitro show stable growth,rapid proliferation,and can be used in tissue engineering.