Abstract
The inability of cultured primary Leydig cells to maintain luteinizing hormone (LH)-responsive testosterone formation in vitro for more than 3–5 days has presented a major challenge in testing trophic effects of regulatory factors or environmental toxicants. Our primary objective was to establish culture conditions sufficient to maintain LH-responsive testosterone formation by Leydig cells for at least a month. When isolated rat adult Leydig cells were cultured in DMEM/F12 and M199 culture medium containing insulin (10μg/ml), PDGFAA (10 ng/ml), lipoprotein (0.25 mg/ml), horse serum (1%) and a submaximal concentration of LH (0.2 ng/ml), the cells retained the ability to produce testosterone in vitro for at least 4 weeks. By using the longer-term culture conditions of this system, we were able to detect suppressive effects on testosterone production by low levels of the toxicant MEHP (mono-(2-ethylhexyl) phthalate), an active metabolite of the plasticizer DEHP, that were not detected by short-term culture.
Original language | English (US) |
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Pages (from-to) | 48-56 |
Number of pages | 9 |
Journal | Molecular and Cellular Endocrinology |
Volume | 476 |
DOIs | |
State | Published - Nov 15 2018 |
Keywords
- Cell culture
- Leydig cells
- Phthalate
- Steroidogenesis
- Testosterone
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Endocrinology