title: Optimizing testosterone production by purified adult rat Leydig cells in vitro.
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.
authors: Department of Population Dynamics, Johns Hopkins School of Hygiene & Public Health, Baltimore, Maryland 21205.
authors: G R
authors: GR
authors: Klinefelter
authors: L L
authors: LL
authors: Ewing
mesh_terms: D000818:Animals; D002469:Cell Separation; D002478:Cells, Cultured; D003911:Dextrans; D007985:Leydig Cells / Q000187:drug effects / Q000378:metabolism*; D008075:Lipoproteins, HDL / Q000494:pharmacology; D008077:Lipoproteins, LDL / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000494:pharmacology; D008297:Male; D008863:Microspheres; D051381:Rats; D011919:Rats, Inbred Strains; D013739:Testosterone / Q000096:biosynthesis*
chemical_list: D003911:Dextrans; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D013739:Testosterone; C052839:Cytodex; D007986:Luteinizing Hormone