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PLoS One ; 7(7): e40469, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22792341

RESUMO

BACKGROUND: The objective of this study was to establish the buffalo mammary epithelial cell line (BuMEC) and characterize its mammary specific functions. METHODOLOGY: Buffalo mammary tissue collected from the slaughter house was processed enzymatically to obtain a heterogenous population of cells containing both epithelial and fibroblasts cells. Epithelial cells were purified by selective trypsinization and were grown in a plastic substratum. The purified mammary epithelial cells (MECs) after several passages were characterized for mammary specific functions by immunocytochemistry, RT-PCR and western blot. PRINCIPAL FINDINGS: The established buffalo mammary epithelial cell line (BuMEC) exhibited epithelial cell characteristics by immunostaining positively with cytokeratin 18 and negatively with vimentin. The BuMEC maintained the characteristics of its functional differentiation by expression of ß-casein, κ-casein, butyrophilin and lactoferrin. BuMEC had normal growth properties and maintained diploid chromosome number (2n = 50) before and after cryopreservation. A spontaneously immortalized buffalo mammary epithelial cell line was established after 20 passages and was continuously subcultured for more than 60 passages without senescence. CONCLUSIONS: We have established a buffalo mammary epithelial cell line that can be used as a model system for studying mammary gland functions.


Assuntos
Búfalos , Células Epiteliais/fisiologia , Glândulas Mamárias Animais/citologia , Animais , Técnicas de Cultura de Células , Proliferação de Células , Forma Celular , Células Cultivadas , Senescência Celular , Cromossomos de Mamíferos/metabolismo , Colágeno/metabolismo , Meios de Cultura , Proteínas do Citoesqueleto/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Feminino , Cariótipo , Proteínas do Leite/genética , Proteínas do Leite/metabolismo , Transcriptoma , beta-Galactosidase/metabolismo
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