鸡贮精腺上皮细胞的分离及原代培养

doi:10.16431/j.cnki.1671-7236.2019.05.009

Abstract

Abstract:

This study was aimed to discuss the stable and reliable method of isolation and primary culture of epithelial cells of sperm storage tubules (SSTs),and provide a cell model for studying the mechanism of chicken sperm storage.The uterovaginal junction of the chicken oviduct was used to culture the epithelial cells of sperm storage tubules with enzyme digestion and explant culture methods.The culture of SSTs cells in hens and the growth of the cells isolated with different cell culture methods was observed.The results showed that after the uterovaginal junction of hen was digested with collagenase or trypsin and then filtered with 100 mesh, the epithelial cells of SSTs could adhere to wall after 24 h,but the cells died after 48-72 h.After the uterovaginal junction of hen was digested with collagenase Ⅺ (0.01 g/mL) and trypsin (0.25%) and then filtered with 100 mesh,there was obvious proliferation at 24 to 48 h,the cells slowly proliferated after 72 h,and then began to die.The epithelial cells of SSTs could be obtained with tissue block culture for 7 days,and the cells could be transferred for 2 to 3 generations.Immunohistochemistry was used to test whether the protein of differentially expressed neurexophilin 1 (NXPH1) of chicken SSTs was located in the primary cells with tissue explant culture.The results showed that NXPH1 expressed in the nuclei and cytoplasm of the cultured cells,indicating that the primary cells with tissue explant culture could be used in subsequent experiments.In conclusion,the epithelial cells of sperm storage tubules obtained with explant culture could provide a cell model for studying the mechanism of sperm storage in hens.

Key words: hens; sperm storage tubules; epithelial cells; cell culture

CLC Number:

Q813.1⁺1

GUO Yuqi, SUN Dongmei, PENG Zhaoyu, JIANG Xunping, HU Xueying, LIU Yun, LIU Guiqiong. Isolation and Primary Culture of Epithelial Cells of Sperm Storage Tubules in Hens[J]. , 2019, 46(5): 1323-1329.

References

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Isolation and Primary Culture of Epithelial Cells of Sperm Storage Tubules in Hens

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