Expression of ACTH-induced corticosteroid biosynthesis in newborn rat adrenocortical cells cultured in serum-free and carrier protein-free medium containing a cholesterol-α-cyclodextrin complex
✍ Scribed by Mohammed Hammami; Gabrielle Maume; Bernard F. Maume
- Publisher
- Springer
- Year
- 1986
- Tongue
- English
- Weight
- 721 KB
- Volume
- 2
- Category
- Article
- ISSN
- 0742-2091
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✦ Synopsis
Newborn rat adrenocortical cells were successfully cultured in a serum-free carrier protein-free medium (SPFM) by using t~cyclodextrin as a cholesterol carrier and have expressed corticosteroid biosynthesis in this medium. A stable inclusion complex of cholesterol-a-cyclodextrin with a molar ratio of almost 1 was obtained for a 5 >( 10 -5 moll1 a-cyclodextrin concentration. Cell cultures incubated with [4 -14 CJ cholesterol-ot-cyclodextrin in SPFM produced, under A CTH stimulation, various 14C labeled steroids with a predominance of corticosterone and 18-hydroxy-11deoxycorticosterone. As measured by gas chromatography and mass spectrometry, the ratio between corticosteroids (21hydroxylated steroids) and 20or-reduced steroids produced in SPFM with cholesterol-a-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-ol-cyclodextrin used in this study is more suitable for corticosteroidogenesis by adrenal cells in culture than a serum-free medium with cholesterol-albumin.