Defined medium for normal adult human prostatic stromal cells |
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Authors: | Donna M Peehl Robert G Sellers Stephen T Wong |
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Institution: | (1) Department of Urology, Stanford University School of Medicine, 94305 Stanford, California |
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Abstract: | Summary Stromal-epithelial interactions are pivotal in many aspects of prostatic biology. A defined culture system is critical for
the investigation of factors that regulate the growth and differentiation of human prostatic stromal cells. We have identified
conditions which promote stromal cell attachment and proliferation in serum-free medium. MCDB 201, originally developed for
the clonal growth of chick embryo fibroblasts, proved to be a superior basal medium of those that we tested. Supplementation
of MCDB 201 with basic fibroblast growth factor (FGF), insulin-like growth factor (IGF), and platelet-derived growth factor
(PDGF) permitted attachment and exponential growth of cells throughout a 7-d period with an initial inoculum as low as 103 cells per well of a 96-well microtiter dish. Using these assay conditions, we subsequently verified that basic FGF and IGF,
but not PDGF, were required for optimal growth. No activity was found for heparin, transferrin, or the androgen R1881. Epidermal
growth factor (EGF) didn’t stimulate growth when added to medium containing basic FGF and IGF, but was moderately stimulatory
when added to basal medium alone. Cholera toxin inhibited growth. This simple and efficient culture medium provides a suitable
assay system for more extensive studies of growth regulation and differentiation of human prostatic stromal cells, and will
provide the basis for future development of a defined medium that supports clonal growth. Characterization of stromal-epithelial
interactions will be facilitated by the use of this defined culture system for stromal cells in conjunction with the serum-free
culture systems previously developed for human prostatic epithelial cells. |
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Keywords: | prostate fibroblast growth factor insulin-like growth factor epidermal growth factor platelet-derived growth factor cholera toxin |
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