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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Published in | Zhonghua nan ke xue Vol. 19; no. 1; p. 29 |
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Main Authors | , , , , , , |
Format | Journal Article |
Language | Chinese |
Published |
China
01.01.2013
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Subjects | |
Online Access | Get more information |
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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 |
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ISSN: | 1009-3591 |