奶牛乳腺上皮细胞原代培养、纯化及鉴定技术研究进展

doi:10.16431/j.cnki.1671-7236.2015.08.027

摘要/Abstract

摘要： 奶牛乳腺上皮细胞不仅具有合成和分泌乳汁的功能,而且在乳腺的先天免疫中扮演着重要角色,对泌乳机制、乳房炎发病机制的研究,以及药物筛选具有重要意义。原代培养的奶牛乳腺上皮细胞适宜建立细胞模型,可作为生理、病理、药理等方面研究的良好介质,解决体内试验周期长、成本高、个体差异大的难题。作者主要从奶牛乳腺上皮细胞原代培养的发展历程、培养技术、纯化技术及鉴定方法等方面的最新研究情况进行综述,以期为奶牛乳腺上皮细胞培养相关研究提供参考。

关键词: 奶牛; 乳腺上皮细胞; 原代培养; 鉴定

Abstract: The bovine mammary epithelial cells not only have the function of synthesis and secretion of milk, but also play an important role in innate immunity system of the mammary gland, and there is great significance of them on studying the mechanism of lactation, mastitis pathogenesis and drug screening.Primary cultured bovine mammary gland epithelial cells are suitable for setting up cell model which can be used as the dielectric for physiological, pathological and pharmacological researching, avoiding the difficulties of in vivo test, such as the long cycle, the high cost, the individual difference, etc.The author summarized the latest researches of cell primary culture in vitro, cultivation technology, purification and identification method in order to provide reference for the studies of bovine mammary epithelial cells culture.

Key words: bovine; mammary epithelial cells; primary culture; identification

中图分类号:

S823

林杰, 王旭荣, 李建喜, 王孝武, 杨志强. 奶牛乳腺上皮细胞原代培养、纯化及鉴定技术研究进展[J]. , 2015, 42(8): 2109-2115.

LIN Jie, WANG Xu-rong, LI Jian-xi, WANG Xiao-wu, YANG Zhi-qiang. Research Progress on Primary Culture,Purification and Identification Technique of Bovine Mammary Epithelial Cells[J]. , 2015, 42(8): 2109-2115.

参考文献

[1] Diego C,Eugenio M,Silvia M,et al.Bovine mammary epithelial cells retain stem-like phenotype in long-term cultures[J].Res Vet Sci,2014,97(2):367-375.
[2] Capuco A V,Choudhary R K,Daniels K M,et al.Bovine mammary stem cells:Cell biology meets production agriculture[J].Animal,2012,6(3):382-393.
[3] Borena B M,Bussche L,Burvenich C,et al.Mammary stem cell research in veterinary science:An update[J].Stem Cells Dev,2013,22(12):1743-1751.
[4] Girolami F,Spalenza V,Manzini L,et al.Constitutive expression of the AHR signaling pathway in a bovine mammary epithelial cell line and modulation by dioxin-like PCB and other AHR ligands[J].Toxicol Lett,2014,232(1):98-105.
[5] Zielniok K,Motyl T,Gajewska M.Functional interactions between 17b-estradiol and progesterone regulate autophagy during acini formation by bovine mammary epithelial cells in 3D cultures[J].Biomed Res Int,2014,2014:1-16.
[6] 王治国.奶牛乳腺上皮细胞的体外培养及应用[D].北京:中国农业科学院,2007.
[7] Lasfargues E Y.Cultivation and behavior in vitro of the normal mammary epithelium of the adult mouse[J].Anat Rec,1957,127(1):117-129.
[8] Ebner K E,Hoover C R,Hageman E C,et al.Cultivation and properties of bovine mammary cell cultures [J].Exp Cell Res,1961,23:373-385.
[9] Medina D,Oborn C J.Growth of preneoplastic mammary epithelial cells in serum-free medium[J].Cancer Res,1980,40(11):3982-3987.
[10] McGrath M.A novel system for mammary epithelial cell culture[J].J Dairy Sci,1987,70(9):1967-1980.
[11] Pantschenko A G,Yang T J.Mitogenic responsiveness of caprine mammary epithelial cells to endocrine and cytokine factors[J].Endocrine,1999,10(2):123-130.
[12] 李震,王英,刘惠莉,等.牛乳腺上皮细胞的分离培养[J].上海农业学报,2000,16(3):25-28.
[13] 赵学军.奶牛乳腺组织体外培养体系的建立及MET对其酪蛋白基因表达的影响[D].杭州:浙江大学,2004.
[14] 彭新荣.牛乳腺上皮细胞的体外分离与培养[D].杨凌:西北农林科技大学,2004.
[15] 弗雷谢尼(英)著.乳腺上皮细胞的培养:动物细胞培养——基础技术指南[M].章静波,徐存拴等译.第4版.北京:科学出版社,2004.
[16] Wellnitz O,Kerr D E.Cryopreserved bovine mammary cells to model epithelial response to infection[J].Vet Immunol Immunopathol,2004,101(3-4):191-202.
[17] Hou X,Li Q,Huang T.Microarray analysis of gene expression profiles in the bovine mammary gland during lactation[J].Sci China Life Sci,2010,53(2):248-256.
[18] Strandberg Y,Gray C,Vuocolo T,et al.Lipopolysaccharide and lipoteichoic acid induce different innate immune responses in bovine mammary epithelial cells[J].Cytokine,2005,31(1):72-86.
[19] Sorg D,Danowski K,Korenkova V,et al.Microfluidic high-throughput RT-qPCR measurements of the immune response of primary bovine mammary epithelial cells cultured from milk to mastitis pathogens[J].Animal,2013,7(5):799-805.
[20] Jedrzejczak M,Szatkowska I.Bovine mammary epithelial cell cultures for the study of mammary gland functions[J].In Vitro Cell Dev Biol Anim,2014,50(9):389-398.
[21] Rauner G,Barash I.Xanthosine administration does not affect the proportion of epithelial stem cells in bovine mammary tissue,but has a latent negative effect on cell proliferation[J].Exp Cell Res,2014,328(1):186-196.
[22] Zbinden C,Stephan R,Johler S,et al.The inflammatory response of primary bovine mammary epithelial cells to Staphylococcus aureus strains is linked to the bacterial phenotype[J].PLoS One,2014,9(1):e87374.
[23] Hagi T,Sasaki K,Aso H,et al.Adhesive properties of predominant bacteria in raw cow's milk to bovine mammary gland epithelial cells[J].Folia Microbiol,2013,58(6):515-522.
[24] Zhao K,Liu H Y,Zhou M M,et al.Establishment and characterization of a lactating bovine mammary epithelial cell model for the study of milk synthesis[J].Cell Biol Int,2010,34(7):717-721.
[25] Zhao K,Liu H Y,Zhou M M,et al.Insulin stimulates glucose uptake via a phosphatidylinositide 3-kinase-linked signaling pathway in bovine mammary epithelial cells[J].J Dairy Sci,2014,97(6):3660-3665.
[26] Wang J L,Gao X J,Zhang L Q,et al.Effect of methionine on lactation ability of dairy cow mammary epithelial cells cultured in vitro[J].J Dairy Sci and Technol,2012,35(1):8-10.
[27] 赵学军.奶牛乳腺组织体外培养体系的建立及MET对其酪蛋白基因a_s1表达的影响[D].杭州:浙江大学,2004.
[28] 刘荣,高学军,李庆章,等.奶牛乳腺上皮细胞核蛋白组双向电泳法的建立[J].中国畜牧兽医,2012,39(2):31-34.
[29] 李文清,王加启,南雪梅,等.奶牛乳腺上皮细胞的不同培养方法的比较及激素和细胞因子对β-酪蛋白mRNA表达的诱导[J].动物营养学报,2014,26(9):2607-2614.
[30] Yang Z,Fu Y,Liu B,et al.Farrerol regulates antimicrobial peptide expression and reduces Staphylococcus aureus internalization into bovine mammary epithelial cells[J].Microb Pathog,2013,65:1-6.
[31] 孙海云,李建国,崔亚利,等.奶牛乳腺上皮细胞培养液中β-酪蛋白提取方法的改进及其对细胞分泌功能的影响[J].中国兽医学报,2014,34(6):982-990.
[32] 罗莹,吴金节,王希春,等.奶牛乳腺上皮细胞培养液中β-酪蛋白提取方法的改进及其对细胞分泌功能的影响[J].畜牧与兽医,2014,46(9):11-14.
[33] 丁旭娜,王凤龙,丁玉林,等.奶牛乳腺上皮细胞和成纤维细胞的体外培养与鉴定[J].黑龙江畜牧兽医,2012,3(5):1-5.
[34] 史琳琳,赵锋,高学军,等.催乳复合物、胰岛素样生长因子-Ⅰ对奶牛乳腺上皮细胞中AKT-1和mTOR表达的作用[J].中国畜牧兽医,2013,40(3):23-27.
[35] 王亨,吴培福,邱昌伟,等.荷斯坦奶牛乳腺上皮细胞的体外培养[J].畜牧与兽医,2007,39(4):41-43.
[36] 多曙光,吴应积,罗奋华,等.牛乳腺上皮细胞的分离培养及其生物学特性[J].动物学研究,2006,27(3):299-305.
[37] Hébert A,Sayasith K,Sénéchal S,et al.Demonstration of intracellular Staphylococcus aureus in bovine mastitis aveolar cells and macrophages isolated from naturally infected cow milk[J].FEMS Microbiol Lett,2000,193(1):57-62.
[38] 崔新洁,王婷,刘秉春,等.牛奶中乳腺上皮细胞的分离培养及鉴定[J].生物技术通报,2013,3:107-113.
[39] 李艳,徐凯,赵国琦.奶牛乳腺上皮细胞的培养与鉴定[J].上海畜牧兽医通讯,2014,1:8-11.
[40] 李吉霞.牛乳腺上皮细胞永生化及其多能性的研究[D].杨凌:西北农林科技大学,2010.
[41] 宋亚男,陈志伟,刘东武.牛乳腺上皮细胞体外培养体系研究进展[J].生命科学仪器,2008,6(9):3-8.
[42] Fu Y,Zhou E,Liu Z,et al.Staphylococcus aureus and Escherichia coli elicit different innate immune responses from bovine mammary epithelial cells[J].Vet Immunol Immunopathol,2013,155(4):245-252.
[43] Zavizion B,Van Duffelen M,Schaeffer W,et al.Establishment and characterization of a bovine mammary epithelial cell line with unique properties[J].In Vitro Cell Dev Biol,1996,32(3):138-148.
[44] Pantschenko A G,Barber M R,Woodcock-Mitchell J,et al.Establishment and characterization of a caprine mammary myoepithelial cell line (CMMyoEC)[J].In Vitro Cell Dev Biol Anim,2000,36:351-356.
[45] 王建民,陈存仙,董飞,等.一种获得山羊乳腺上皮细胞单细胞克隆的方法[P].中国:201310430440,2014.
[46] 克莱尔·怀斯(英)主编.段恩奎,王莉等译.上皮细胞培养指南[M].北京:科学出版社,2005.
[47] 曹琪娜.采用自制培养基培养奶牛乳腺上皮细胞可行性研究[D].呼和浩特:内蒙古农业大学,2012.
[48] Han Z Y,Mu T,Yang Z.Methionine protects against hyperthermia-induced cell injury in cultured bovine mammary epithelial cells[J].Cell Stress Chaperones,2015,20(1):109-120.
[49] Tong J J,Ye L,Liu R,et al.Effect of semen vaccariae and taraxacu mogono on cell adhesion of bovine mammary epithelial cells[J].Journal of Integrative Agriculture,2012,11(12):2043-2050.
[50] Sun H Y,Li J G,Cui Y L,et al.Improvement of extraction method for beta-casein in dairy cow mammary epithelial cells culture and the influence on the cell secretion function[J].Chinese Journal of Veterinary Science,2014,34(6):982-985.