Summary
This report demonstrates that routine variations in cell culture conditions dramatically affect the amount of vitamin D3 metabolites to which cultured cells have access. Increasing the concentration of a metabolite in the medium increases the amount of the metabolite in the cell compartment. Increasing the volume of medium in the culture dishes (while maintaining a constant metabolite concentration) also increases the amount of metabolite in the cell compartment. Moreover, daily changes of the medium containing fresh metabolite increase the amount of the metabolite in the cell compartment as well. These variables may explain the inability of different laboratories to duplicate dose-response curves.
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References
Henry HL (1981) Insulin permits parathyroid hormone stimulation of 1,25-dihydroxyvitamin D3 production in cultured kidney cells. Endocrinology 108:733–735
Henry HL (1981) 25(OH)D3 metabolism in kidney cell cultures: Lack of a direct effect of estradiol. Am J Physiol 240:E119-E124
Turner RT, Bottemiller BL, Howard GA, Baylink DJ (1980)In vitro metabolism of 25-hydroxyvitamin D3 by isolated rat kidney cells. Proc Natl Acad Sci USA 77:1537–1540
Turner RT, Puzas JE, Forte MD, Lester GE, Gray TK, Howard GA, Baylink DJ (1980)In vitro synthesis of 1-alpha-25-dihydroxycholecalciferol and 24,25-dihydroxycholecalciferol by isolated calvarial cells. Proc Natl Acad Sci USA 77:5720–5724
Treschel U, Bonjour J-P, Fleisch H (1979) Regulation of the metabolism of 25-hydroxyvitamin D3 in primary cultures of chick kidney cells, J Clin Invest 64:206–217
Treschel U, Bonjour J-P, Fleisch H (1979) Regulation of 25-hydroxylases in cultured kidney cells. In:Vitamin D, basic research and its clinical application. Walter De Gruyter & Co., Berlin, New York, p 597–604
Howard GA, Turner RT, Sherrard DJ, Baylink DJ (1981) Human bone cells in culture metabolize 25-hydroxyvitamin D3 to 1,25-dihydroxyvitamin D3 and 24,25-dihydroxyvitamin D3. J Biol Chem 256:7738–7740
Puzas JE, Drivdahl RH, Howard GA, Baylink DJ (1981) Endogenous inhibitor of bone cell proliferation. Proc Soc Exp Biol Med 166:113–122
Puzas JE, Turner RT, Howard GA, Brand JS (1981) Studies on the metabolism of vitamin D3 by chick bone cells. In: Norman AW, Schaefer K, Herrath DV, Grigoliet HG, (eds)Vitamin D: Chemical, biochemical and clinical endocrinology of calcium metabolism. Walter de Gruyter, New York, p 13–16
Majeska RJ, Rodan GA (1981) The effect of 1,25(OH)2D3 on alkaline phosphatase in osteoblastic osteosarcoma cells. J Biol Chem 257:3362–3365
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Puzas, J.E., Brand, J.S. In vitro use of vitamin D3 metabolites: Culture conditions determine cell uptake. Calcif Tissue Int 37, 474–477 (1985). https://doi.org/10.1007/BF02557829
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DOI: https://doi.org/10.1007/BF02557829