Characterization of the effects of defined, multidimensional culture conditions on conditionally reprogrammed primary human prostate cells
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Lucas Tricoli1,4, Aisha Naeem1, Erika Parasido1, John P. Mikhaiel1, Muhammad Umer Choudhry1, Deborah L. Berry1, Iman A. Abdelgawad2, Richard J. Lee3, Adam S. Feldman3, Chukwuemeka Ihemelandu1, Maria Avantaggiati1, Deepak Kumar4, Stephen Byers1, Rosa Gallagher5, Julia Wulfkuhle5, Emanuel Petricoin5, Olga Rodriguez1,6 and Chris Albanese1,6
1Department of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University Medical Center, Washington, DC, USA
2National Cancer Institute of Egypt, Cairo, Egypt
3Massachusetts General Hospital Cancer Center, Boston, MA, USA
4Julius L. Chambers Biomedical/Biotechnology Research Institute, North Carolina Central University, Durham, NC, USA
5Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, VA, USA
6Preclinical Imaging Research Laboratory, Georgetown University Medical Center, Washington, DC, USA
Chris Albanese, email: [email protected]
Keywords: prostate; cancer; primary tissue; reprogrammed cells; androgen
Received: October 26, 2017 Accepted: November 02, 2017 Published: December 18, 2017
The inability to propagate human prostate epithelial cells indefinitely has historically presented a serious impediment to prostate cancer research. The conditionally reprogrammed cell (CRC) approach uses the combination of irradiated J2 mouse fibroblasts and a Rho kinase inhibitor such as Y27632 to support the continuous culture of cells derived from most epithelial tissues, including the prostate. Due to their rapid establishment and overall ease of use, CRCs are now widely used in a variety of basic and preclinical settings. In addition, CRCs were successfully used to clinically treat respiratory papillomatosis. Although both normal and tumor-derived prostate CRCs have been used to study the basic biology of prostate cancer and to test new therapies, certain limitations exist. We have previously reported that prostate CRCs form functional prostate glands when implanted under the mouse renal capsule. However in conventional culture, the prostate CRCs exist in an adult stem-like, transient amplifying state and consequently do not adequately recapitulate several important features of a differentiated prostate epithelium. To address these limitations, we previously described a transwell dish-based model that supported the culturing of prostate CRCs and the collection of cells and cell extracts for molecular and genetic analyses. Using normal and tumor-derived prostate CRCs, we describe the combined effects of the multi-dimensional transwell platform and defined culture media on prostate cellular proliferation, differentiation and signaling.
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