裸鼹鼠成纤维细胞原代培养方法的建立

doi:10.3969/j.issn.1674-5817.2013.06.015

实验动物与比较医学 ›› 2013, Vol. 33 ›› Issue (6): 481-486.DOI: 10.3969/j.issn.1674-5817.2013.06.015

裸鼹鼠成纤维细胞原代培养方法的建立

林丽芳, 赵善民, 肖邦, 汤球, 张璐, 袁子彦, 孙智远, 于琛琳, 蔡丽萍, 徐晨, 孙伟, 崔淑芳

第二军医大学实验动物中心, 上海 200433

收稿日期:2013-07-15 出版日期:2013-12-25 发布日期:2013-12-25
作者简介:林丽芳(1986-), 女, 助教。E-mail: linlifang2012@126.com。
基金资助:
上海市科技发展基金项目(No. 12140900200)与 (No.13140900400)联合资助

Establishment of Primary Culture Method of Fibroblast Derived from Naked Mole Rats

LIN Li-fang, ZHAO Shan-min, XIAO Bang, TANG Qiu, ZHANG Lu, YUAN Zi-yan, SUN Zhi-yuan, YU Chen-lin, CAI Li-ping, XU Chen, SUN Wei, CUI Shu-fang

Laboratory Animal Center, Second Military Medical University,Shanghai 200433, China

Received:2013-07-15 Online:2013-12-25 Published:2013-12-25

摘要/Abstract

摘要： 目的建立裸鼹鼠成纤维细胞的原代培养方法,研究裸鼹鼠成纤维细胞的生长特性。方法结合传统的消化培养法和组织培养法进行裸鼹鼠和小鼠成纤维细胞原代培养,CCK-8(cell counting kit)细胞计数比较研究两种物种成纤维细胞的生长速率和细胞周期。以不同浓度接种裸鼹鼠肝脏成纤维细胞,CCK-8计数细胞24 h、72 h、96 h、144 h细胞密度。结果结合消化培养法和组织培养法两种细胞原代培养方法,裸鼹鼠及小鼠成纤维细胞的生长状态良好。裸鼹鼠成纤维细胞的增殖速率显著高于小鼠成纤维细胞,处于对数生长期的细胞比例与小鼠比较,显著较高。以1.5×10⁴/ml 以上的浓度接种裸鼹鼠成纤维细胞能使细胞在两天内进入细胞生长对数期。结论结合消化培养法和组织培养法的原代培养方法能提高细胞原代培养的效率,裸鼹鼠成纤维细胞的增殖能力显著高于小鼠。

关键词: 裸鼹鼠成纤维细胞, 小鼠成纤维细胞, 原代培养方法, 细胞增殖速率

Abstract: Objective To established the primary culture method of fibroblasts derived from naked mole rat, and research the growth characteristics of the cells. Methods The fibroblasts derived from naked mole rat and mouse were cultured by method which combined traditional enzymes digestion with explant culture.A comparative study between fibroblasts derived from two kinds of species on growth rate and cell cycle was conducted. Fibroblasts derived from liver of naked mole rats were inoculated with density gradients, then counted the cells after 24 h, 72 h, 96 h, 144 h. Results Fibroblasts derived from naked mole rat and mouse had a good growth behavior with the primary culture method combining enzymes digestion and tissue block culture. Compared to mouse, fibroblasts derived from naked mole rats had higher proportion in the logarithmic growth phase and proliferation rate significantly. Fibroblasts derived from naked mole rat can reach logarithmic phase within two days, seeded with the concentration of 1.5×10⁴/ml. Conclusion The method which combined enzymes digestion with tissue block culture improved efficiency of cell culture. Moreover, the proliferation ability of fibroblasts derived from naked mole rat were significantly higher than mice.

Key words: Naked mole rat, Mouse, Fibroblasts, Primary culture, Cell proliferation rate

林丽芳, 赵善民, 肖邦, 汤球, 张璐, 袁子彦, 孙智远, 于琛琳, 蔡丽萍, 徐晨, 孙伟, 崔淑芳. 裸鼹鼠成纤维细胞原代培养方法的建立[J]. 实验动物与比较医学, 2013, 33(6): 481-486.

LIN Li-fang, ZHAO Shan-min, XIAO Bang, TANG Qiu, ZHANG Lu, YUAN Zi-yan, SUN Zhi-yuan, YU Chen-lin, CAI Li-ping, XU Chen, SUN Wei, CUI Shu-fang. Establishment of Primary Culture Method of Fibroblast Derived from Naked Mole Rats[J]. Laboratory Animal and Comparative Medicine, 2013, 33(6): 481-486.

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裸鼹鼠成纤维细胞原代培养方法的建立

Establishment of Primary Culture Method of Fibroblast Derived from Naked Mole Rats

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