现代商品仔猪传代肾细胞株的培育与应用研究

doi:10.16431/j.cnki.1671-7236.2017.02.039

Abstract

Abstract:

Cell line was an important carrier for the isolation and culture of animal viruses.In the research, modern commercial piglet kidney was taken as the raw material in order to cultivate a new cell line for isolating and culturing animal viruses. By trypsin digestion and differential velocity adherent combination method,porcine kidney epithelial cells were separated and purified,and then continuously subcultured in vitro. The results showed that a new cell strain named SDPK-D had been serially passaged for 90 generations.The doubling time of SDPK-D cell strain F33 and F83 were 40.9 and 32.7 h,respectively;While the cell viability were 97.55% and 98.86%,the cell adherent rate at the 8 h were 91.67% and 97.06%,respectively. Obvious cytopathic effect appeared after inoculating with PRV,SIV and PEDV positive samples. For the first time, a new cell strain from modern pig named SDPK-D was successfully cultivated,which was sensitive for several animal viruses.It could provid a new selection for the isolation and culture of virus.

Key words: piglet kidney cell; primary culture; subculture; cell strain

CLC Number:

Q813.1

HU Hong-hui, WU Jing-bo, HUANG Jian-qiang, NAN Wen-jin, YU Wen-tao. Study on Cultivation Research and Preliminary Application of Modern Commercial Piglet Kidney Cell Strain[J]. , 2017, 44(2): 586-593.

References

[1] Freshney R I. Culture of Animal Cell:A Manual of Basic Technique[M].6th ed.New Jersey:Wiley-Blackwell,2010.
[2] 殷震,刘景华.动物病毒学[M].第2版.北京:科学出版社,1997.
[3] 司徒镇强,吴军正.细胞培养[M].北京:世界图书出版公司,2001.
[4] 李群,钱钟,范娟,等.动物细胞悬浮培养技术在兽用疫苗生产领域中的应用[J].中国畜牧兽医,2013,40(6):236-240.
[5] 梅建国,庄金秋,王金良,等.动物细胞大规模培养技术[J].中国生物工程杂志,2012,32(7):127-132.
[6] Kim H,Lee Y K,Kang S C,et al. Recent vaccine technology in industrial animals[J].Clin Exp Vaccine Res,2016,5(1):12-18.
[7] Zuñiga S,Pascual-Iglesias A,Sanchez C M,et al.Virulence factors in porcine coronaviruses and vaccine design[J].Virus Res, 2016,226(6):142-151.
[8] Crotty S,Pipkin M E.In vivo RNAi screens:Concepts and applications[J].Trends Immunol,2015,36(5):315-322.
[9] 段盼盼.Vero细胞无血清培养基的研制及其在流感病毒培养中的应用[D].北京:北京协和医学院,2015.
[10] 周峰,杜贤,董长垣,等.原代肾上皮细胞改良培养方法[J].中国组织工程研究与临床康,2009,13(11):2101-2104.
[11] 陈娴,刘畅,张晴晴,等.关于鼠肾小管上皮细胞原代培养的对比探究[J].世界最新医学信息文摘,2015,19(15):13-16.
[12] 王东,吴雄飞,金锡御.大鼠肾小管上皮细胞的原代培养及传代[J].中华实验外科杂志,1999,3(2):179-180.
[13] 王献伟,卢晟盛,卢克焕.新生广西巴马小型猪肾成纤维细胞体外培养体系的建立[J].中国兽医学报,2011,31(10):1526-1532.
[14] Puck T T,Marcus P I.A rapid method for viable cell titration and clone production with HeLa cells in tissue culture:The use of X-irradiated cells to supply conditioning factors[J].Proc Natl Acad Sci USA,1955,41(7):432-437.
[15] 阮小华.合作猪肾纤维细胞系的构建及生物学鉴定[D].兰州:西北民族大学,2010.
[16] Liu H,Lv P,Zhu X,et al.In vitro development of porcine transgenic nuclear-transferred embryos derived from newborn Guangxi Bama mini-pig kidney fibroblasts[J]. In Vitro Cell Dev Biol Anim,2014,50(9):811-821.
[17] Guerin L F,Guerin M M.Susceptibility of pig kidney tissue cultures to certain viruses[J].Proc Soc Exp Biol Med,1957,96(2):322-323.
[18] Sellers R F,Stewart D L.Factors affecting the formation of plaques by the virus of food-and-mouth disease on pig kidney monolayer tissue cultures[J].Arch Virol,1960,9(5):594-605.
[19] Abelseth M K.Propagation of rabies virus in pig kidney cell culture[J].Can Vet J,1964,5(4):84-87.
[20] 姜春霞,马广鹏,姜艳平,等.猪传染性胃肠炎病毒LJ-12株的分离与鉴定[J].畜牧与兽医,2013,45(3):47-50.
[21] Kumar R,Barman N N,Khatoon E,et al.Molecular characterization of E2 glycoprotein of classical swine fever virus:Adaptation and propagation in porcine kidney cells[J].In Vitro Cell Dev Biol Anim,2015,51(5):441-446.
[22] 戚丹英,刘福安.幼猪肾(BPKV)传代细胞系病毒感染范围[J].畜牧兽医科技,1985,1:23-27.
[23] 毛凝,郑敏,黄梅清,等.猪肾细胞感染猪伪狂犬病毒后的细胞观察与分析[J].台湾农业探索,2013,4(2):70-72.

Study on Cultivation Research and Preliminary Application of Modern Commercial Piglet Kidney Cell Strain

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