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Cell Stem Cell. 2008 Jan 10;2(1):90-102. doi: 10.1016/j.stem.2007.10.002.

Lentiviral transduction of mammary stem cells for analysis of gene function during development and cancer.

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1
Department of Anatomy, University of California, San Francisco, 513 Parnassus Avenue, San Francisco, CA 94143-0452, USA.

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.

PMID:
18371425
PMCID:
PMC2276651
DOI:
10.1016/j.stem.2007.10.002
[Indexed for MEDLINE]
Free PMC Article

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