Isolation, culture, and identification of duck intestinal epithelial cells and oxidative stress model constructed
Intestinal epithelial cells (IECs) not only have an absorption function but also act as a physical barrier between the body and the intestinal bacterial flora. Damage to IECs leads to the breakdown of this barrier and has negative effects on animal health. Intestinal epithelial damage is frequently associated with long-term acute stress, such as increased temperature and new stress management models. The intestinal epithelial damage caused by environmental stress has been linked to oxidative stress. Until now, the effects of intestinal epithelial antioxidant activity from feed additives and treatments could be tested in ducks only in vivo because of the lack of in vitro cell culture systems. In this study, we describe our protocol for the easy isolation and culture of IECs from the small intestine of duck embryos. Immunofluorescence was used for the cytological identification of IECs. In addition, IEC marker genes (IAP and CDH1) could also be detected in cultured cells. And cell status assessments were performed, and cell proliferation viability was analyzed by CCK-8 assay. Furthermore, we constructed an oxidative stress model to be used to research the oxidative stress response mechanism, and drugs acting on the cell signal transduction pathway. In conclusion, we have developed an effective and rapid protocol for obtaining duck primary IECs and constructed an oxidative stress model. These IECs exhibit features consistent with epithelial cells and could be used to explore the physiological mechanisms of oxidative stress ex vivo.
KeywordsIntestinal epithelial cells Primary cultivation Identification Duck
This work was supported by grants from the National Natural Science Foundation (Grant numbers: 31702157), the Hubei Provincial scientific and technological innovation special project (Grant Numbers: 2017ABA140) and China Agriculture Research System (Grant Numbers: CARS-42-47), and Hubei Academy of Agricultural Sciences Younger Top-Notch Talent Program (Grant Numbers: Q2018021). Animal slaughter followed the Ethics Committee of the Institute of Hubei Academy of Agricultural sciences.
Compliance with ethical standards
All animal experiments were approved and carried out in accordance with the Animal Care and Use Committee of Hubei Academy of Agricultural Sciences and in accordance with the principles and guidelines of the National Institutes of Health.
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