Effects of cholesterol and cAMP on progesterone production in cultured luteal cells isolated from pseudopregnant cat ovaries
Yükleniyor...
Tarih
2009
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Elsevier Science Bv
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
The present study was designed to incubate luteal cells isolated from pseudopregnant cats and to investigate the effects of cholesterol and cAMP on luteal progesterone production. Corpora lutea were collected from the cats on days 10 and 15 of pseudopregnancy. Luteal cells were isolated from the ovaries by collagenase digestion. Steroidogenic luteal cells were stained for 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity. Cells (2 x 10(4)) staining positive for 3 beta-HSD were cultured for up to 7 days. The cells were treated with 22(R)-hydroxycholesterol (22R-HC) and dibutyryl cyclic AMP (dbcAMP) on days 1, 3 and 7. Treatment of cells with 22R-HC resulted in a dose-dependent increase (p < 0.001) in progesterone production. When 22R-HC was used at a concentration of 10 mu g/ml, it resulted in 2.7- and 5.1-fold increases in progesterone production on days 3 and 5, respectively. When the dose was doubled (20 mu g/ml), treated cells produced four times more progesterone on days 3 and 7, and three times more on day 5. By day 7, progesterone production increased up to 9.1 times more than the control. Incubation of cells with both concentrations of dbcAMP (0.1 mM and 1 mM) resulted in significant stimulations of progesterone on days 5 and 7 (p < 0.001). However, on day 3, only higher doses of dbcAMP (1 mM) resulted in significant stimulation (p < 0.05). Progesterone production was increased up to 2- and 2.9-fold of the control when cells were treated with lower concentration of dbcAMP (0.1 mM) on days 5 and 7. respectively. Incubation of cells with 1 mM concentrations of dbcAMP induced a 3.2-fold increase on day 5 and a 5-fold increase on day 7. In conclusion, a successful incubation was performed for long-life culturing of luteal cells collected from pseudopregnant cats. The method works well and allows for optimal growth and development of cells in the culture. The present study also demonstrated that incubating cat luteal cells with 22R-HC and dbcAMP induces a significant increase in luteal progesterone synthesis. (C) 2008 Elsevier B.V. All rights reserved.
Açıklama
Anahtar Kelimeler
Cell culture, Cats, Pseudopregnancy, Progesterone, 22R-HC, dbcAMP
Kaynak
Animal Reproduction Science
WoS Q Değeri
Q1
Scopus Q Değeri
Q1
Cilt
115
Sayı
1-4
Künye
closedAccess
