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

doi:10.3969/j.issn.1674-5817.2013.06.015

Abstract

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

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.

References

[1] Buffenstein R.The naked mole-rat:a new long-living model for human aging research[J].J Gerontol A Biol Sci Med Sci, 2005, 60(11):1369-1377.
[2] Sherman PW, Jarvis JUM.Extraordinary life spans of naked mole-rats (Heterocephalus glaber)[J]. J of Zool, 2002, 258(3):307-311.
[3] O’Connor TP, Lee A, Jarvis JU, et al. Prolonged longevity in naked mole-rats:age-related changes in metabolism,body composition and gastrointestinal function[J].Comp Biochem Physiol A Mol Integr Physiol, 2002, 133(3):835-842.
[4] Fukagawa NK, Bandini LG, et al.Effect of age on body composition and resting metabolic rate[J].Am J Physiol, 1990, 59(2):E233-E238.
[5] 孙伟, 汤球, 赵善民, 等. 裸鼹鼠人工饲养繁殖初步研究[J]. 实验动物与比较医学, 2013, 33(4):296-300.
[6] 袁子彦, 赵懿宁, 张璐, 等. 裸鼹鼠肺脏显微结构与超微结构观察[J]. 实验动物与比较医学, 2013, 33(5):378-382.
[7] 袁子彦, 赵懿宁, 张璐, 等. 裸鼹鼠肝脏显微结构与超微结构观察[J]. 实验动物与比较医学, 2013, 33(5):373-377.
[8] 赵善民, 赵懿宁, 汤球, 等. 裸鼹鼠胸腺、脾脏及淋巴结解剖学、组织学与超微结构研究[J]. 实验动物与比较医学, 2013, 33(5):395-399.
[9] 赵善民, 崔淑芳. 新型实验动物裸鼹鼠生物学特性的研究进展[J]. 实验动物与比较医学, 2013, 33(5):400-405.
[10] 张璐, 赵懿宁, 袁子彦, 等. 裸鼹鼠与C57BL/6J小鼠肾脏结构及超微结构比较[J]. 实验动物与比较医学, 2013, 33(5):388-394.
[11] Liang S, Mele J, Wu Y, et al.Resistance to experimental tumorigenesis in cells of a long-lived mammal, the naked mole-rat (Heterocephalus-glaber)[J]. Aging Cell, 2010, 9(4):626-635.
[12] Salmon AB, Sadighi Akha AA, Buffenstein R, et al.Fibroblasts from naked mole-rats are resistant to multiple forms of cell injury, but sensitive to peroxide, UV Light, and ER Stress[J]. J Gerontol A Biol Sci Med Sci, 2008, 63(3):232-241.
[13] Seluanov A, Hine C, Azpurua J, et al.Hypersensitivity to contact inhibition provides a clue to cancer resistance of naked mole-rat[J]. PNAS, 2009, 106(46):19352-19357.
[14] Bruce D, Sharry L, Thomas L, et al.Factors influencing metabolic rate in naked mole-rats (Heteroce phalus glaber)[J]. Physiol Behav, 1999, 66(3):447-459.
[15] McNab BK. The metabolism of fossorial rodents: A study of convergence[J]. Ecology, 1966, 47(5):712-732.