Comparison of marker protein expression in benign prostatic hyperplasiain vivoandin vitro

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Abstract
Objective To use multiple immunofluorescence to compare the in vivo and in vitro expression of tissue‐specific proteins in BPH. Materials and methods Pure populations of prostate epithelial and stromal cells were produced using standard methods. Serum‐free media for epithelial cells were compared. Co‐localization of proteins was compared in frozen‐tissue sections and cultured cells by simultaneous multiple immunofluorescence, and recorded using a high‐resolution charge‐coupled device camera. Results In contrast to the other serum‐free media tested, epithelial cells grew without squamous differentiation or vacuolation in prostate epithelial growth medium (PrEGM, Clonetics, BioWhittaker UK Ltd., Berks, UK). These cells were predominantly of a basal phenotype, with some cells showing a luminal phenotype. Most of the stromal cells had features of myofibroblasts, but smooth muscle cells and fibroblasts also were present. Conclusion PrEGM is a commercially available serum‐free medium in which primary cultures of prostate epithelial cells can be propagated reproducibly. This study provides a comprehensive description of tissue‐specific protein expression in BPH in vivo and in vitro. The use of simultaneous multiple immunofluorescence to study co‐localization has resulted in a more precise definition of phenotype than has previously been possible, thereby establishing the relevance of the in vitro model system BPH.