Neoplastic transformation of porcine mammary epithelial cells in vitro and XXXX formation in vivo.

@entity1 @entity1707 @entity675 @entity0 @entity986 @entity5 @entity19 @entity42327 @entity876 @entity7596 @entity9617
BACKGROUND: The mammary glands of @entity1707 share many functional and morphological similarities with the breasts of @entity1 , raising the potential of their utility for research into the mechanisms underlying normal mammary function and breast carcinogenesis. Here we sought to establish a model for the efficient manipulation and transformation of porcine mammary epithelial cells (pMEC) in vitro and @entity5 growth in vivo. METHODS: We utilized a vector encoding the red florescent protein tdTomato to transduce populations of pMEC from Yorkshire -Hampshire crossbred female @entity1707 in vitro and in vivo. Populations of primary pMEC were then separated by FACS using markers to distinguish epithelial cells ( @entity675 -) from stromal cells ( @entity675 +), with or without further enrichment for basal and luminal progenitor cells ( @entity9617 +). These separated pMEC populations were transduced by lentivirus encoding murine polyomavirus T antigens (Tag) and tdTomato and engrafted to orthotopic or ectopic sites in immunodeficient NOD.Cg-Prkdc (scid) @entity42327 (tm1Wjl) /SzJ (NSG) @entity19 . RESULTS: We demonstrated that lentivirus effectively transduces pMEC in vitro and in vivo. We further established that lentivirus can be used for oncogenic-transformation of pMEC ex vivo for generating @entity5 in vivo. Oncogenic transformation was confirmed in vitro by anchorage-independent growth, increased cell proliferation, and expression of @entity986 , @entity7596 and @entity876 alongside decreased phosphorylation of Rb. Moreover, Tag-transformed @entity675 - and @entity675 - @entity9617   +   pMECs developed site-specific @entity5 of differing histopathologies in vivo. CONCLUSIONS: Herein we establish a model for the transduction and oncogenic transformation of pMEC. This is the first report describing a porcine model of mammary epithelial cell tumorigenesis that can be applied to the study of @entity1 @entity0 .
