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.