富勒醇对基于悬滴培养的脂肪间充质干细胞成骨分化的影响  

Effects of fullerol on the osteogenic differentiation of adipose-derived mesenchymal stem cells based on hanging drop culture

作　　者：唐艳婷 孙岩峰[2] 李小军 周瑾[3] 郝彤[3] 李俊杰[3] 张琪[4] 

机构地区：[1]锦州医科大学,辽宁锦州121001 [2]武警总医院儿科,北京100039 [3]军事医学科学院基础医学研究所前沿交叉学科研究室,北京100850 [4]武警总医院口腔颌面外科,北京100039

出　　处：《解放军医学杂志》2016年第8期624-628,共5页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(31370975)~~

摘　　要：目的探讨富勒醇对基于悬滴培养的大鼠脂肪间充质干细胞(r ADSCs)向成骨细胞分化的影响。方法将rADSCs通过悬滴培养3d形成大鼠脂肪间充质干细胞球,将细胞球用胰酶消化分散成单细胞,对获得的单细胞进行二维贴壁培养24h,然后更换为成骨诱导培养基培养,其中未添加富勒醇组为对照组,添加1.0μmol/L富勒醇组为实验组,诱导分化14d及21d后利用茜素红染色和实时定量PCR检测脂肪干细胞球来源的单细胞向成骨细胞分化的能力。结果悬滴培养3d的rADSCs可形成大小均一的微球结构,经胰酶消化分散可获得细胞球来源的单细胞。与对照组相比,在成骨诱导培养基中添加1.0μmol/L富勒醇可使所获单细胞形成更多的矿化的钙结节,且相关成骨基因Runx2、OCN、ColⅠ的表达增强。结论富勒醇可以显著增强基于悬滴培养获得的rADSCs的成骨分化,有助于提高其成骨诱导的效率。Objective To investigate the effect of fullerol on the osteogenic differentiation of rat adipose-derived mesenchymal stem cells（r ADSCs） based on hanging drop culture. Methods r ADSC spheres were formed by hanging drop culture for 3 days, then the spheres were dissociated to single cell by trypsin（called r ADSC sphere-derived cells）. The r ADSC spherederived cells were cultured in 2-D adherent cell cultures for 24 hours, then the ordinary culture medium was replaced by osteogenic induction medium（osteogenic medium, OM）. OM without fullerol served as control group（CM）; OM with 1.0μmol/L fullerol was used as the experimental group（OM＋Ful1.0μmol/L）. Two groups were induced to differentiation for 14 d and 21 d, using two methods of alizarin red staining and quantitative real-time PCR（q PCR） to detecte the ability of r ADSC sphere-derived cells differentiating into osteoblasts. Results r ADSCs could form a uniform size microspheres structure through hanging drop culture for 3 days; r ADSC sphere-derived cells were obtained by dissipation of trypsin. Compared with OM group, OM＋Ful1.0μmol/L can make r ADSC sphere-derived cells form more mineralized calcium nodules, and enhance the expression of the relevant osteogenic gene Runx2, OCN and Col I. Conclusion Fullerol can significantly enhance the osteogenic differentiation of r ADSCs based on hanging drop culture, and it is helpful to improve the efficiency of osteogenic induction of r ADSCs.

关 键 词：间质干细胞 细胞分化 富勒醇 悬滴培养 

分 类 号：R318.5[医药卫生—生物医学工程]