Characterization of primary human mammary epithelial cells isolated and propagated by conditional reprogrammed cell culture
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Liting Jin1,2,*, Ying Qu2,*, Liliana J. Gomez2, Stacey Chung2, Bingchen Han2, Bowen Gao2, Yong Yue3, Yiping Gong1, Xuefeng Liu4, Farin Amersi2, Catherine Dang2, Armando E. Giuliano2 and Xiaojiang Cui2
1Department of Breast Surgery, Hubei Cancer Hospital, Wuhan, 430079, China
2Department of Surgery, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA
3Department of Radiation, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical Center, Los Angeles, CA 90048, USA
4Department of Pathology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, DC 20057, USA
*These authors contributed equally to the work
Xiaojiang Cui, email: Xiaojiang.Cui@cshs.org
Keywords: conditional reprogramming; estrogen receptor; mammary epithelial cells; luminal cells; myoepithelial cells
Received: April 07, 2017 Accepted: October 30, 2017 Published: December 22, 2017
Purpose: Conditional reprogramming methods allow for the inexhaustible in vitro proliferation of primary epithelial cells from human tissue specimens. This methodology has the potential to enhance the utility of primary cell culture as a model for mammary gland research. However, few studies have systematically characterized this method in generating in vitro normal human mammary epithelial cell models.
Results: We show that cells derived from fresh normal breast tissues can be propagated and exhibit heterogeneous morphologic features. The cultures are composed of CK18, desmoglein 3, and CK19-positive luminal cells and vimentin, p63, and CK14-positive myoepithelial cells, suggesting the maintenance of in vivo heterogeneity. In addition, the cultures contain subpopulations with different CD49f and EpCAM expression profiles. When grown in 3D conditions, cells self-organize into distinct structures that express either luminal or basal cell markers. Among these structures, CK8-positive cells enclosing a lumen are capable of differentiation into milk-producing cells in the presence of lactogenic stimulus. Furthermore, our short-term cultures retain the expression of ERα, as well as its ability to respond to estrogen stimulation.
Materials and Methods: We have investigated conditionally reprogrammed normal epithelial cells in terms of cell type heterogeneity, cellular marker expression, and structural arrangement in two-dimensional (2D) and three-dimensional (3D) systems.
Conclusions: The conditional reprogramming methodology allows generation of a heterogeneous culture from normal human mammary tissue in vitro. We believe that this cell culture model will provide a valuable tool to study mammary cell function and malignant transformation.
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