Expression of ACTH-induced corticosteroid biosynthesis in newborn rat adrenocortical cells cultured in serum-free and carrier protein-free medium containing a cholesterol-alpha-cyclodextrin complex.
Newborn rat adrenocortical cells were successfully cultured in a serum-free carrier protein-free medium (SPFM) by using alpha-cyclodextrin as a cholesterol carrier and have expressed corticosteroid biosynthesis in this medium. A stable inclusion complex of cholesterol-alpha-cyclodextrin with a molar ratio of almost 1 was obtained for a 5 X 10(-5) mol/1 alpha-cyclodextrin concentration. Cell cultures incubated with [4-14C] cholesterol-alpha-cyclodextrin in SPFM produced, under ACTH stimulation, various 14C labeled steroids with a predominance of corticosterone and 18-hydroxy-11-deoxycorticosterone. As measured by gas chromatography and mass spectrometry, the ratio between corticosteroids (21-hydroxylated steroids) and 20 alpha-reduced steroids produced in SPFM with cholesterol-alpha-cyclodextrin was equal to 1.8. This corresponds to a value of 3.6 times higher than that found in the serum-free medium with cholesterol-albumin. Consequently, the chemically defined SPFM with cholesterol-alpha-cyclodextrin used in this study is more suitable for corticosteroidogenesis by adrenal cells in culture than a serum-free medium with cholesterol-albumin.
Hammami
M
M
Laboratoire de Biochimie des Interactions Cellulaires, Faculté de Sciences, Dijon, France.
Maume
G
G
Maume
B F
BF
D000302:Adrenal Cortex / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D000305:Adrenal Cortex Hormones / Q000096:biosynthesis*; D000324:Adrenocorticotropic Hormone / Q000494:pharmacology*; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism / Q000493:pharmacokinetics*; D003470:Culture Media / Q000494:pharmacology; D003505:Cyclodextrins / Q000378:metabolism / Q000493:pharmacokinetics*; D003912:Dextrins / Q000493:pharmacokinetics*; D051381:Rats; D013213:Starch / Q000493:pharmacokinetics*; D047391:alpha-Cyclodextrins
D000305:Adrenal Cortex Hormones; D003470:Culture Media; D003505:Cyclodextrins; D003912:Dextrins; D047391:alpha-Cyclodextrins; C061207:cholesterol-alpha-cyclodextrin; D000324:Adrenocorticotropic Hormone; D013213:Starch; D002784:Cholesterol