PMID- 24831228 OWN - NLM STAT- MEDLINE DCOM- 20150129 LR - 20211021 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 9 IP - 5 DP - 2014 TI - Conditionally reprogrammed normal and transformed mouse mammary epithelial cells display a progenitor-cell-like phenotype. PG - e97666 LID - 10.1371/journal.pone.0097666 [doi] LID - e97666 AB - Mammary epithelial (ME) cells cultured under conventional conditions senesce after several passages. Here, we demonstrate that mouse ME cells isolated from normal mammary glands or from mouse mammary tumor virus (MMTV)-Neu-induced mammary tumors, can be cultured indefinitely as conditionally reprogrammed cells (CRCs) on irradiated fibroblasts in the presence of the Rho kinase inhibitor Y-27632. Cell surface progenitor-associated markers are rapidly induced in normal mouse ME-CRCs relative to ME cells. However, the expression of certain mammary progenitor subpopulations, such as CD49f+ ESA+ CD44+, drops significantly in later passages. Nevertheless, mouse ME-CRCs grown in a three-dimensional extracellular matrix gave rise to mammary acinar structures. ME-CRCs isolated from MMTV-Neu transgenic mouse mammary tumors express high levels of HER2/neu, as well as tumor-initiating cell markers, such as CD44+, CD49f+, and ESA+ (EpCam). These patterns of expression are sustained in later CRC passages. Early and late passage ME-CRCs from MMTV-Neu tumors that were implanted in the mammary fat pads of syngeneic or nude mice developed vascular tumors that metastasized within 6 weeks of transplantation. Importantly, the histopathology of these tumors was indistinguishable from that of the parental tumors that develop in the MMTV-Neu mice. Application of the CRC system to mouse mammary epithelial cells provides an attractive model system to study the genetics and phenotype of normal and transformed mouse epithelium in a defined culture environment and in vivo transplant studies. FAU - Saenz, Francisco R AU - Saenz FR AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Ory, Virginie AU - Ory V AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - AlOtaiby, Maram AU - AlOtaiby M AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Rosenfield, Sonia AU - Rosenfield S AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Furlong, Mary AU - Furlong M AD - Department of Pathology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Cavalli, Luciane R AU - Cavalli LR AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Johnson, Michael D AU - Johnson MD AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Liu, Xuefeng AU - Liu X AD - Department of Pathology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Schlegel, Richard AU - Schlegel R AD - Department of Pathology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Wellstein, Anton AU - Wellstein A AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. FAU - Riegel, Anna T AU - Riegel AT AD - Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, United States of America. LA - eng GR - P30CA051008/CA/NCI NIH HHS/United States GR - R33 CA177466/CA/NCI NIH HHS/United States GR - CA113477/CA/NCI NIH HHS/United States GR - R01 CA113477/CA/NCI NIH HHS/United States GR - CA18052/CA/NCI NIH HHS/United States GR - P30 CA051008/CA/NCI NIH HHS/United States GR - R21 CA180524/CA/NCI NIH HHS/United States GR - CA177466/CA/NCI NIH HHS/United States GR - T32 CA009686/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20140515 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Amides) RN - 0 (Drug Combinations) RN - 0 (Enzyme Inhibitors) RN - 0 (Laminin) RN - 0 (Proteoglycans) RN - 0 (Pyridines) RN - 119978-18-6 (matrigel) RN - 138381-45-0 (Y 27632) RN - 9007-34-5 (Collagen) SB - IM MH - Amides/chemistry MH - Animals MH - Cell Transformation, Neoplastic/genetics MH - Cells, Cultured MH - Coculture Techniques MH - Collagen/chemistry MH - Comparative Genomic Hybridization MH - Drug Combinations MH - Enzyme Inhibitors/chemistry MH - Epithelial Cells/*cytology MH - Epithelial-Mesenchymal Transition MH - Extracellular Matrix/metabolism MH - Female MH - Fibroblasts/metabolism MH - Laminin/chemistry MH - Mammary Glands, Animal/*cytology MH - Mammary Neoplasms, Experimental/*pathology MH - Mammary Tumor Virus, Mouse/*metabolism MH - Mice MH - Mice, Transgenic MH - Microscopy, Confocal MH - Phenotype MH - Proteoglycans/chemistry MH - Pyridines/chemistry MH - Stem Cells/cytology PMC - PMC4022745 COIS- Competing Interests: Georgetown University has a patent pending on the cell reprogramming technology (Immortalizing Epithelial Cells and Methods of Use, Application No. 13/885,078) and has licensed the technology to a new biotechnology company, Propagenix (www.propagenix.com). Georgetown University also has founding equity in Propagenix. RS and XL are Professors at Georgetown University who are consultants to Propagenix and RS has founding equity in the company. This does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. EDAT- 2014/05/17 06:00 MHDA- 2015/01/30 06:00 PMCR- 2014/05/15 CRDT- 2014/05/17 06:00 PHST- 2014/03/12 00:00 [received] PHST- 2014/04/10 00:00 [accepted] PHST- 2014/05/17 06:00 [entrez] PHST- 2014/05/17 06:00 [pubmed] PHST- 2015/01/30 06:00 [medline] PHST- 2014/05/15 00:00 [pmc-release] AID - PONE-D-14-11305 [pii] AID - 10.1371/journal.pone.0097666 [doi] PST - epublish SO - PLoS One. 2014 May 15;9(5):e97666. doi: 10.1371/journal.pone.0097666. eCollection 2014.