ISSN:
1573-6822
Keywords:
ACTH
;
adrenocortical cell culture
;
cholesterol-α-cyclodextrin complex
;
corticosteroid biosynthesis
;
newborn rat
;
serum-free protein-free medium
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
,
Medicine
Notes:
Abstract Newborn rat adrenocortical cells were successfully cultured in a serum free carrier protein free medium (SPFM) by using α-cyclodextrin as a cholesterol carrier and have expressed corticosteroid biosynthesis in this medium. A stable inclusion complex of cholesterol-α-cyclodextrin with a molar ratio of almost 1 was obtained for a 5 × 10−5 mol/1 α-cyclodextrin concentration. Cell cultures incubated with [4-14C] cholesterol-α-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 20α-reduced steroids produced in SPFM with cholesterol-α-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-α-cyclodextrin used in this study is more suitable for corticosteroidogenesis by adrenal cells in culture than a serum free medium with cholesterol-albumin.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00117706
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