上海交通大学学报（医学版）

›› 2011, Vol. 31 ›› Issue (6): 733-.doi: 10.3969/j.issn.1674-8115.2011.06.012

• 论著（基础研究） • 上一篇    下一篇

新型非病毒纳米基因载体PEI-β-CyD对软骨细胞和骨髓间充质干细胞转染有效性评估

童海骏1, 黄金刚1, 汤谷平2, 戴尅戎1,3, 张晓玲1,3   

  1. 1.上海交通大学医学院/中国科学院上海生命科学研究院 健康科学研究所 骨科细胞与分子生物学实验室, 上海 200025； 2.浙江大学化学生物和药物研究所, 杭州 310028； 3.上海交通大学 医学院附属第九人民医院骨科 上海市骨科内植物重点实验室, 上海 200011
  • 出版日期:2011-06-28 发布日期:2011-06-27
  • 通讯作者: 张晓玲, 电子信箱: xlzhang@sibs.ac.cn。
  • 作者简介:童海骏（1985—）, 男, 硕士；电子信箱: korean.moon@yahoo.com.cn。
  • 基金资助:

    国家科技部国际合作项目（2011DFA30790）；国家自然科学基金国际合作项目（30811120440）；国家自然科学基金（30871435）；上海市科委国际合作项目（09540703600）；上海教委重点学科建设基金（J50206）

Evaluation of transfection efficiency of novel non-viral nano gene vector PEI-β-CyD on chondrocytes and bone marrowderived mesenchymal stem cells

TONG Hai-jun1, HUANG Jin-gang1, TANG Gu-ping2, DAI Ke-rong1,3, ZHANG Xiao-ling1,3   

  1. 1.Laboratory of Orthopedical Cellular and Molecular Biology, Institute of Health Sciences, Shanghai Jiaotong University School of Medicine &|Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China；2.Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou 310028, China；3.Shanghai Key Laboratory of Orthopaedic Implant, Department of Orthopaedics, the Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China
  • Online:2011-06-28 Published:2011-06-27
  • Supported by:

    Foundation of the Ministry of Science and Technology of China, S2011GR0323；National Natural Science Foundation of China, 30811120440, 30871435；Foundation of Shanghai Science and Technology Committee, 09540703600；Foundation of Shanghai Education Committee, J50206

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摘要：

目的 评估新型非病毒纳米基因载体聚乙烯亚胺-β-环糊精（PEI-β-CyD）对软骨细胞系C5.18和骨髓间充质干细胞系C3H10T1/2转染的有效性。方法 体外培养软骨细胞系C5.18和骨髓间充质干细胞系C3H10T1/2，MTT法观察并比较PEI-β-CyD和相对分子质量为25 000的聚乙烯亚胺（PEI25KDa）对C5.18细胞和C3H10T1/2细胞的细胞毒性。在不同的N/P（载体有效氮含量/外源基因有效磷含量）复合条件下，利用PEI-β-CyD和PEI25KDa分别转染C5.18细胞和C3H10T1/2细胞，倒置荧光显微镜结合流式细胞仪分析转染效率。结果 PEI-βCyD对C5.18细胞和C3H10T1/2细胞的细胞毒性均小于PEI25KDa。PEI-β-CyD与PEI25KDa对C5.18细胞的转染效率比较，差异无统计学意义（P>0.05）；PEI-βCyD对C3H10T1/2细胞的转染效率显著高于PEI25KDa（P<0.05）。结论 PEI-β-CyD对于软骨细胞系C5.18及骨髓间充质干细胞系C3H10T1/2是一种低毒有效的非病毒纳米基因载体，在软骨组织工程和细胞治疗研究及应用领域具有良好的开发前景。

关键词: 非病毒纳米基因载体, PEI-β-CyD, 细胞毒性, 转染效率

Abstract:

Objective To evaluate the transfection efficiency of novel non-viral nano gene vector polyethyleneimine-β-cyclodextrin (PEI-β-CyD) on chondrocyte cell line C5.18 and bone marrow-derived mesenchymal stem cell line C3H10T1/2. Methods Chondrocyte cell line C5.18 and bone marrow-derived mesenchymal stem cell line C3H10T1/2 were cultured in vitro, and the cytotoxicity of PEI-β-CyD and PEI25KDa (polyethyleneimine of 25 000 in molecular weight) on C5.18 cells and C3H10T1/2 cells was observed and compared by MTT method. C5.18 cells and C3H10T1/2 cells were transfected with PEI-β-CyD and PEI25KDa respectively under different ratios of effective nitrogen content in vectors to effective phosphorus content in exogenous gene (N/P), and the transfection efficiency was analysed by inverted fluorescent microscopy and flow cytometry. Results The cytotoxicity of PEI-β-CyD was significantly lower than that of PEI25KDa to both C5.18 cells and C3H10T1/2 cells. For C5.18 cells, there was no significant difference in transfection efficiency between PEI-β-CyD and PEI25KDa (P>0.05), and for C3H10T1/2 cells, transfection efficiency of PEI-β-CyD was significantly higher than that of PEI25KDa (P<0.05). Conclusion PEI-β-CyD is an ideal non-viral nano gene vector with low cytotoxicity and favorable transfection efficiency for chondrocyte cell line C5.18 and bone marrow-derived mesenchymal stem cell line C3H10T1/2, which is promising in the research and application of cartilage tissue engineering and cell therapy.

Key words: non-viral nano gene vector, PEI-β-CyD, cytotoxicity, transfection efficiency