ADENOVIRAL-MEDIATED GENE-TRANSFER INTO PRIMARY HUMAN AND MOUSE MAMMARY EPITHELIAL-CELLS IN-VITRO AND IN-VIVO

Primary mammary epithelial cells from both the human and mouse mammary glands can be genetically altered under a variety of situations using the replication-defective adenoviral vector containing a marker gene encoding the E, coli beta-galactosidase. Primary human and mouse mamma ry epithelial cells in monolayer culture and in three-dimensional coll agen gel culture systems were transduced by adenovector at high effici ency. Successful gene transfer was also accomplished in situ and in vi vo. In the mouse mammary gland, anatomically restricted gene transfer and expression was demonstrated by microinjection of adenoviral vector directly into the main duct of the mammary gland. Injection of adenov iral vector directly into the human mammary tissues from reduction mam moplasty specimens, into the mouse mammary gland-free fat pad containi ng the previously transplanted dissociated human mammary epithelial ce lls, and intratumorally into the human breast cancer xenografts in nud e mice, all resulted in successful gene transfer to human mammary epit helial cells. High efficiency introduction of genetic material into pr imary mammary epithelial cells is important in the study of mammary ca rcinogenesis and potentially for gene therapy of human breast cancer.