Inhibition of proliferation of contaminating fibroblasts by D-valine in cultures of smooth muscle cells from human myometrium.
Replacement of L-valine with D-valine in a standard culture medium can selectively inhibit fibroblast proliferation. The aim of the present study was to investigate whether human myometrial cells cultured with D-valine instead of L-valine can survive and express their characteristics. Cultured cells (95-98%) maintain expression of the intermediate filament desmin, which is the specific marker for mature muscle cells. By transmission electron microscopy, the cells showed the general morphology of smooth muscle cells in culture. Oxytocin in serum-free culture medium at 37 degrees C (5 min) caused a concentration-dependent increase in cellular Na and total Ca, and a decrease in K content as determined by X-ray microanalysis. The percentage of cells cultured with D-valine responding to oxytocin stimulation was larger than that of cells cultured with L-valine, suggesting less contamination of smooth muscle cells by fibroblasts in the presence of D-valine. As shown by measurements with fura-2, D-valine-cultured cells retained the characteristic increase in intracellular free Ca2+ ions after oxytocin stimulation.
Hongpaisan
J
J
Department of Medical Cell Biology, Univeristy of Uppsala, Sweden. jarin@codon.nih.gov
D002118:Calcium / Q000378:metabolism; D002455:Cell Division; D002478:Cells, Cultured; D003470:Culture Media; D016895:Culture Media, Serum-Free; D003893:Desmin / Q000096:biosynthesis; D004577:Electron Probe Microanalysis; D005260:Female; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron; D009215:Myometrium / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D010121:Oxytocin / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D013237:Stereoisomerism; D014633:Valine / Q000494:pharmacology*
D003470:Culture Media; D016895:Culture Media, Serum-Free; D003893:Desmin; D010121:Oxytocin; D012964:Sodium; D014633:Valine; D011188:Potassium; D002118:Calcium