Hypoxia reduces the proliferation and occludin expression of primary sertoli cells

To investigate the effect of hypoxia on the proliferation and occludin expression of primary rat Sertoli We constructed a primary Sertoli cell system by two-step enzymatic digestion in 18 -22 days old Wistar rats and identified it by oil red O and immunofluorescence methods. We randomly divided the...

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Bibliographic Details
Published inZhonghua nan ke xue Vol. 19; no. 1; p. 29
Main Authors Hao, Wei-Yu, Shao, Cui-Hua, Feng, You-Liang, Hu, Jian-Ting, Li, Qiang, Wang, Hong-Qiang, Wang, Pei-Tao
Format Journal Article
LanguageChinese
Published China 01.01.2013
Subjects
Animals
Cell Hypoxia
Cell Proliferation
Cells, Cultured
Male
Occludin - metabolism
Rats
Rats, Wistar
Sertoli Cells - metabolism
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Summary:To investigate the effect of hypoxia on the proliferation and occludin expression of primary rat Sertoli We constructed a primary Sertoli cell system by two-step enzymatic digestion in 18 -22 days old Wistar rats and identified it by oil red O and immunofluorescence methods. We randomly divided the Sertoli cells into five groups to be cultured in oxygen at the concentrations of 20%, 15%, 10%, 5% and 1%, respectively, for 6, 12, 24, 48 and 72 hours. We detected the proliferation of the Sertoli cells by CCK-8 assay, determined the expression of occludin by Western blot, and analyzed the differences among the five groups. Oil red O staining revealed red lipid droplets in the cytoplasm of the Sertoli cells, and immunofluorescence showed the positive expression of the FasL protein, with the purity of Sertoli cells over 95% in vitro. Compared with the 20% normoxic group, the proliferation of the Sertoli cells was gradually reduced in the 15% and 10% hypoxia groups, and significantly declined in the 5% and 1% groups
ISSN:1009-3591