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  • Title: Optimizing testosterone production by purified adult rat Leydig cells in vitro.
    Author: Klinefelter GR, Ewing LL.
    Journal: In Vitro Cell Dev Biol; 1988 Jun; 24(6):545-9. PubMed ID: 2455704.
    Abstract:
    We sought to establish conditions that increased the duration of testosterone production by fully differentiated adult rat Leydig cells in primary culture. A freshly isolated suspension of highly purified adult rat Leydig cells produced 83 ng testosterone/10(6) Leydig cells.h-1 when incubated in Medium 199 in a 1.5 ml microfuge tube with shaking for 3 h with a maximally stimulating concentration of ovine luteinizing hormone (LH). Unfortunately, adult rat Leydig cells that were allowed to attach only to a plastic culture dish flattened out, and testosterone production diminished rapidly. Leydig cells in Dulbecco's modified Eagles' medium-Ham's F12 (1:1; vol/vol) containing Cytodex 3 beads pre-equilibrated in culture medium containing fetal bovine serum attached to the beads and remained viable, but produced only 30 ng testosterone/10(6) Leydig cells.h-1 when incubated for 24 h with similar stimulation. Leydig cells similarly cultured and maximally stimulated with LH, responded to bovine lipoproteins (less than 1.222 g/ml) producing 105 ng of testosterone/10(6) Leydig cells.h-1 when incubated with 1 mg/ml bovine lipoprotein. Therefore, lipoproteins maintain the steroidogenic capacity of purified adult rat Leydig cells in primary culture for 24 h.
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