Development and validation of immortalized bovine mammary epithelial cell line as an in vitro model for the study of mammary gland functions

被引:3
|
作者
Li, Ji-Xia [1 ]
Said, Abdelrahman [2 ,3 ]
Ge, Xiu-Guo [1 ]
Wang, Wenxiu [4 ]
Zhang, Yong [5 ]
Jin, Tianming [1 ]
机构
[1] Tianjin Agr Univ, Coll Anim Sci & Vet Med, Tianjin 300384, Peoples R China
[2] Natl Res Ctr, Parasitol & Anim Dis Dept, Giza, Egypt
[3] Univ Saskatchewan, Vaccine & Infect Dis Org, 120 Vet Rd, Saskatoon, SK S7N 5E3, Canada
[4] Shandong Binzhou Anim Sci & Vet Med Acad, Binzhou 256600, Shandong, Peoples R China
[5] Northwest A&F Univ, Coll Vet Med, Inst Biotechnol, Yangling 712100, Shaanxi, Peoples R China
基金
中国国家自然科学基金; 美国国家科学基金会;
关键词
Bovine mammary epithelial cell; Immortalization; Milk protein expression; TELOMERASE REVERSE-TRANSCRIPTASE; HERPESVIRUS TYPE-1 EHV-1; CATALYTIC SUBUNIT; REPLICATIVE SENESCENCE; MAC-T; GROWTH-HORMONE; MESSENGER-RNA; LIFE-SPAN; ESTABLISHMENT; EXPRESSION;
D O I
10.1007/s10616-017-0114-3
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
This study aimed to develop a bovine mammary epithelial (BME) cell line model, which provides a possibility to determine functional properties of the bovine mammary gland. The primary cell culture was derived from bovine mammary gland tissues and processed enzymatically to obtain cell colonies with epithelial-like morphology. The cultures of BME cells were purified and optimally cultured at 37 A degrees C in DMEM/F12 medium supplemented with 10% fetal bovine serum. The BME cells were identified as epithelial cell line by the evaluating the expression of keratin-18 using immunofluorescence staining. A novel gene expression system strongly enhances the expression of telomerase, has been used to immortalize BME cell line termed hTBME cell line. Interestingly, telomerase remained active even after over 60 passages of hTBME cell line, required for immortalization of BME cells. In addition, the hTBME cell line was continuously subcultured with a spontaneous epithelial-like morphology, with a great proliferation activity, and without evidence of apoptotic and necrotic effects. Further characterization showed that hTBME cell line can be continuously propagated in culture with constant chromosomal features and without tumorigenic properties. Finally, established hTBME cell line was evaluated for mammary gland specific functions. Our results demonstrated that the hTBME cell line was able to retain functional-morphological structure, and functional differentiation by expression of beta (beta)-casein as in the bovine mammary gland in vivo. Taken together, our findings suggest that the established hTBME cell line can serve as a valuable tool for the study of bovine mammary gland functions.
引用
收藏
页码:67 / 82
页数:16
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