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) |
|---|---|
| Journal | Molecular and Cellular Endocrinology |
| DOIs | |
| State | Accepted/In press - Jan 1 2018 |
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Keywords
- Cell culture
- Leydig cells
- Phthalate
- Steroidogenesis
- Testosterone
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Endocrinology
Cite this
Long -term maintenance of luteinizing Hormone–Responsive testosterone formation by primary rat Leydig cells in vitro. / Wang, Yiyan; Huang, Shengsong; Wang, Zhao; Chen, Fenfen; Chen, Panpan; Zhao, Xingxing; Lin, Han; Ge, Renshan; Zirkin, Barry R; Chen, Haolin.
In: Molecular and Cellular Endocrinology, 01.01.2018.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Long -term maintenance of luteinizing Hormone–Responsive testosterone formation by primary rat Leydig cells in vitro
AU - Wang, Yiyan
AU - Huang, Shengsong
AU - Wang, Zhao
AU - Chen, Fenfen
AU - Chen, Panpan
AU - Zhao, Xingxing
AU - Lin, Han
AU - Ge, Renshan
AU - Zirkin, Barry R
AU - Chen, Haolin
PY - 2018/1/1
Y1 - 2018/1/1
N2 - 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.
AB - 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.
KW - Cell culture
KW - Leydig cells
KW - Phthalate
KW - Steroidogenesis
KW - Testosterone
UR - http://www.scopus.com/inward/record.url?scp=85046171184&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85046171184&partnerID=8YFLogxK
U2 - 10.1016/j.mce.2018.04.008
DO - 10.1016/j.mce.2018.04.008
M3 - Article
C2 - 29702242
AN - SCOPUS:85046171184
JO - Molecular and Cellular Endocrinology
JF - Molecular and Cellular Endocrinology
SN - 0303-7207
ER -