title: Lentiviral transduction of mammary stem cells for analysis of gene function during development and cancer.
abstract: The mouse mammary gland is the only epithelial organ capable of complete regeneration upon orthotopic transplantation, making it ideally suited for in vivo gene function studies through viral-mediated gene delivery. A hurdle that has challenged the widespread adoption of this technique has been the inability to transduce mammary stem cells effectively. We have overcome this limitation by infecting total primary mammary epithelial cells in suspension with high-titer lentiviruses. Transduced cells gave rise to all major cell types of the mammary gland and were capable of clonal outgrowth and functional differentiation in serial transplants. To demonstrate that this method is a valuable alternative to developing transgenic animals, we used lentiviral-mediated Wnt-1 overexpression to replicate MMTV-Wnt-1 mammary phenotypes and used a dominant-negative Xenopus Suppressor of Hairless to reveal a requirement for Notch signaling during ductal morphogenesis. Importantly, this method is also applicable to transduction of cells from other tissues.
authors: Department of Anatomy, University of California, San Francisco, 513 Parnassus Avenue, San Francisco, CA 94143-0452, USA.
authors: Bryan E
authors: BE
authors: Welm
authors: Gerrit J P
authors: GJ
authors: Dijkgraaf
authors: Anita S
authors: AS
authors: Bledau
authors: Alana L
authors: AL
authors: Welm
authors: Zena
authors: Z
authors: Werb
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004847:Epithelial Cells / Q000502:physiology; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D008321:Mammary Glands, Animal / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D008325:Mammary Neoplasms, Experimental / Q000235:genetics / Q000473:pathology; D008324:Mammary Tumor Virus, Mouse; D051379:Mice; D009024:Morphogenesis; D051880:Receptors, Notch / Q000502:physiology; D015398:Signal Transduction; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics; D014161:Transduction, Genetic; D051155:Wnt1 Protein / Q000096:biosynthesis / Q000235:genetics
chemical_list: D051880:Receptors, Notch; D014157:Transcription Factors; D051155:Wnt1 Protein