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Main Title Maintenance of Testosterone Production by Purified Adult Rat Leydig Cells for 3 Days In vitro.
Author Klinefelter, G. R. ; Ewing., L. L. ;
CORP Author Johns Hopkins Univ., Baltimore, MD. School of Hygiene and Public Health.;Health Effects Research Lab., Research Triangle Park, NC.
Publisher c1989
Year Published 1989
Report Number EPA/600/J-89/481;
Stock Number PB91-117572
Additional Subjects Testosterone ; Biosynthesis ; Rats ; In vitro analysis ; Cytochromes ; Reprints ; Leydig cells ; LH ; Cultured cells ; Lipoproteins
Library Call Number Additional Info Location Last
NTIS  PB91-117572 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 8p
Abstract Using a preparation of highly purified, adult rat Leydig cells and conditions of culture which was found to optimize testosterone production during 24 h, attempts were made to maintain optimal testosterone production for 3 d. Leydig cells cultured on Cytodex 3 beads at 19% O2 in Dulbecco's modified Eagle's medium-Ham's nutrient mixture F12 (1:1; vol/vol) containing 0.5 mg/ml total bovine lipoproteins (<1.222g/ml) with maximal luteinizing hormone (LH) stimulation failed to maintain a constant amount of testosterone for 3 d. These cells did however secrete a similar amount of total delta 4-3-ketosteroids on each of the 3 culture d, indicating that their viability was preserved. The predominance of progesterone and 170H-progesterone relative to the amount of androstenedione found on Days 2 and 3 suggested that the activity of the cytochrome P450C17-hydroxylase-C17,20-lyase enzyme in the smooth endoplasmic reticulum was diminished when Leydig cells were maintained in our primary culture for longer than 24 h. Decreasing the oxygen tension of the cultures from 19 to 5%, and decreasing the concentration of LH used to stimulate the Leydig cells from 100 to 0.1 ng/ml, were necessary to achieve maintenance of testosterone secretion without accumulation of other delta 4-3-ketosteroids during a 3-d period. Cells cultured in this fashion were still able to respond to maximal LH stimulation during Day 3, producing as much testosterone as if cultured for 24 h on Day 1 at 19% O2 with 100 ng/ml LH stimulation. (Copyright (c) 1989 Tissue Culture Association, Inc.)
Supplementary Notes Pub. in In vitro Cellular and Developmental Biology, v25 n3 pt1 p283-288 Mar 89. Sponsored by Health Effects Research Lab., Research Triangle Park, NC.
NTIS Title Notes Journal article.
Title Annotations Reprint: Maintenance of Testosterone Production by Purified Adult Rat Leydig Cells for 3 Days In vitro.
Category Codes 57F; 57B
NTIS Prices PC A02/MF A01
Primary Description 600/10
Document Type NT
Cataloging Source NTIS/MT
Control Number 101124249
Origin NTIS
Type CAT