Long -term maintenance of luteinizing Hormone–Responsive testosterone formation by primary rat Leydig cells in vitro

Publication date: Available online 24 April 2018
Source:Molecular and Cellular Endocrinology
Author(s): Yiyan Wang, Shengsong Huang, Zhao Wang, Fenfen Chen, Panpan Chen, Xingxing Zhao, Han Lin, Renshan Ge, Barry Zirkin, Haolin Chen
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.



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