兔骨髓间充质干细胞的分离培养及其生物学性状的研究.PDF

中国骨伤 2004 年 5 月第 17 卷第 5 期 　China J Ort hop Trauma ,May . 2004 ,Vol . 17 ,No . 5 ·263 · ·论著 · 兔骨髓间充质干细胞的分离培养及其 生物学性状的研究 1 1 2 1 1 1 1 杨自权 ,卫小春 ,郝一勇 ,焦强 ,丁娟 ,李鹏翠 ,陈崇伟 ( 1 山西医科大学第二医院骨科 ,山西 　太原 　03000 1 ; 2 太原钢铁总公司医院骨科) 　　摘要 　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 ,平均倍增时间为 32 h ,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 。体外培养的兔骨髓间充 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 。 关键词 　骨髓 ; 　间充质干细胞 ; 　细胞培养 ; 　生物学性状 Study of the isolation culture and biological characteristics of rabbit bone marrowderived mesenchymal stem cells in vitro 　Yang Ziquan , Wei X iaochun , Hao Yiy ong , J IA O Qiang , DIN G J uan , L I Pengcui , CH EN Chunw ei . Dep art ment of Ort hop aedics , The S econ d Hosp ital of S hanx i M edical University ( S hanx i ) Taiy uan , 03000 1, China Abstract Objective :To establish a met hod to isolate and culture rabbit bone marrowderived mesenchymal stem cells (BM SCs) and investigate t heir biological characteristics. Methods :Bone marrow aspirate of New Zealand white rabbit was density gradient centrifuged and cultured in plastic culture bottles. The cells were ex amined by invert microscop e ,scanning and transmission electron microscop e . The growt h curve was drawn . Re sults :95 % cells were alive after density gradient centrifugation . BM SCs had a similar spindlelike morp hology . The average time for cell doubling was 32 hours. The proliferation abilit y declined wit h time of t he culture . Conclusion :The combination of density gradient centrifugation and adhere culture is an effective met hod to isolate BM SCs from bone marrow aspirate .