Phenotypic and functional characterization of two bovine mammary epithelial cell lines in 2D and 3D models
2016
AREVALO TURRUBIARTE, Magdalena | Perruchot, Marie-Hélène | Finot, Laurence | Mayeur, Frederique | Dessauge, Frederic
Immortalized bovine mammary epithelial cells (BME-UV1) and immortalized bovine mammary alveolar cells (MAC-T) have been extensively used as in vitro cell models to understand milk production in dairy cows. Precise knowledge about their phenotype and performance remains however unknown. This study aims to characterize MAC-T and BME-UV1 profiles when cultured in 2D adherent, 3D adherent (Matrigel™) and 3D no adherent (ultra-low attachment ULA) supports. MAC-T and BME-UV1 were compared according to their proliferation capacities and to specific cell surface markers CD24, CD326 (EpCAM), CD10 and integrin CD49f (α-6). Cytokeratin (CK14 and CK19), signal transducer and activator of transcription 5 (STAT5) and other proteins (Occludin and cadherin-1) were analyzed. BME-UV1 in ULA support expressed higher CD49f marker. A different intensity of CD49 staining allowed the discrimination between the 2 cell lines in adherent condition. CD10, EpCAM and CK19 expressions show that BME-UV1 cells have luminal capacity, while MAC-T has a myoepithelial profile with a high expression of CK14. BME-UV1 cells possess a closer committed progenitor profile due to their higher expression in ALDH and EpCAM. We observed that BME-UV1 cells have a better capacity to form spherical structures - mammospheres - in Matrigel ™ than MAC-T, which was confirmed based on the higher mammospheres area. In ULA condition, BME-UV1 proliferated over the 6 days of culture. Taken together, our results clearly confirm the BME-UV1 luminal profile and MAC-T ductal/myoepithelial-like phenotype.
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