Metabolism of 4-14C-Testosterone by Serially Subcultured Human Skin Fibroblasts1

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Abstract
The metabolism of testosterone by cultured human fibroblasts derived from the skin of the prepuce, labia majora, abdomen, and the deltoid region of the arm have been studied. Monolayer cultures were incubated with 4-14C-testosterone for 12, 24, 48, 72, and 96 hours in the absence of a NADPH-generating system and the conversion products were recovered from the medium. These studies revealed that testosterone was metabolized very rapidly by cell cultures derived from sex-skins, namely, the foreskin and the labial skin; whereas, cell cultures derived from non-sex skins, the abdominal and the deltoid skin, metabolized testosterone slowly, thus providing proof that the rate of metabolism of testosterone by a serially subcultured cell strain was a function of the anatomical site of the primary explant. The sequence of appearance of conversion products of testosterone deduced from these studies was: androstenedione, androstanedione, androsterone, epiandrosterone and dihydrotestosterone and finally the 3α- and the 3β-androstanediols. Three other polar metabolites could not be identified. Dihydrotestosterone was shown to be a relatively minor conversion product in the absence of an NADPH-generating system in the medium. Furthermore, the 5β-reduction products, such as 5β-androstanedione, 5β-dihydrotestosterone, and etiocholanolone were absent in the medium.

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