Serial culture of single adult human prostatic epithelial cells in serum-free medium containing low calcium and a new growth factor from bovine brain.
Primary cultures of epithelial cells from human prostate acini proliferate in defined medium. However, the limited availability of human tissue and the lack of knowledge of the conditions required for clonal growth and serial culture of epithelial cells have limited progress in the study of human prostate cell biology. Here we report conditions that permit the proliferation of single epithelial cells from normal, benign hyperplastic, and carcinomatous prostate through three to four serial passages, which represents at least seven to nine cumulative doublings of the cell populations. Primary cultures were prepared from prostatic acini. Monolayers resulting from the outgrowth of epithelial cells from acini were harvested and dissociated into suspensions of single cells which gave rise to discrete colonies in subsequent culture. The requirements for successful serial culture were (a) a low calcium concentration, (b) the presence of a growth factor that is concentrated in bovine neural tissue, (c), detachment of the epithelial cells with collagenase, and (d) harvest of cells before the cell concentration reached 6000 cells/cm2 of culture surface. Suspensions of single cells were successfully stored between subcultures in 10% dimethylsulfoxide with 5% fetal bovine serum and revived after storage for up to 2 months in liquid nitrogen.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media; D004847:Epithelial Cells; D006133:Growth Substances / Q000302:isolation & purification*; D006728:Hormones / Q000494:pharmacology; D006801:Humans; D007031:Hypothalamus / Q000032:analysis; D008297:Male; D003012:Microbial Collagenase / Q000378:metabolism; D011467:Prostate / Q000166:cytology*; D014357:Trypsin / Q000378:metabolism
D003470:Culture Media; D006133:Growth Substances; D006728:Hormones; D014357:Trypsin; D003012:Microbial Collagenase; D002118:Calcium