ISOLATION AND CHARACTERIZATION OF A NOVEL HUMAN PROSTATIC STROMAL CELL-CULTURE - DUK50

A novel human prostatic stromal cell culture, designated DuK50, has be en passed in vitro > 12 mo. Tissue cultures were obtained from materia l harvested within a normal region of a radical prostatectomy specimen . These monolayers exhibited normal fibroblastic characteristics with each cell having a flattened, elongated appearance. Karyotypic analysi s revealed a normal, male 46, XE; chromosomal content with no numerica l or structural abnormalities. DNA analysis using a Cell Analysis Syst ems Image Analyzer confirmed a euploid DNA content (7.9 Dg DNA). Cellu lar markers for verification of stromal cell type were performed by im munohistochemical techniques. DuK50 stained positive for vimentin and fibronectin. Immunostains for epithelial cytokeratins and prostate-spe cific antigen were negative, which ruled out contamination with prosta tic epithelial cells. Negative immunostaining with desmin monoclonal a ntibody and light staining with smooth muscle actin alpha is consisten t with the staining pattern of myofibroblasts. Response to various and rogens, measured by a microculture tetrazolium assay technique, reveal ed a significant growth stimulation of DuK50. Soft agar invasiveness a ssays and tumorigenicity studies in nude mice were negative. DuK50 exh ibits a rapid doubling time with excellent plating efficiency, thrives in a readily available media supplemented with fetal bovine serum, an d passes with routine trypsin protocols. The availability of this pros tatic stromal cell culture may facilitate studies on this cell type's role in modulation, drug and steroid metabolism, and stromal-epithelia l interactions in the prostate.