Isolation and characterization of cultured chicken oviduct epithelial cells and in vitro validation of constructed ovalbumin promoter in these cells

• Published in: Animal bioscience 2021, Vol.34 (8), p.1321-1330
• Main Authors: Yang, Hyeon, Lee, Bo Ram, Lee, Hwi-Cheul, Jung, Sun Keun, Kim, Ji-Youn, No, Jingu, Shanmugam, Sureshkumar, Jo, Yong Jin, Lee, Haesun, Hwang, Seongsoo, Byun, Sung June
• Format: Article
• Language: Korean
• ISSN: 2765-0189; 2765-0235

Objective: Transgenic hens hold a great promise to produce various valuable proteins. Through virus transduction into stage X embryo, the transgene expression under the control of constructed chicken ovalbumin promoters has been successfully achieved. However, a validation system that can evaluate differently developed ovalbumin promoters in in vitro, remains to be developed. Methods: In the present study, chicken oviduct epithelial cells (cOECs) were isolated from oviduct tissue and shortly cultured with keratinocyte complete medium supplemented with chicken serum. The isolated cells were characterized with immunofluorescence, western blot, and flow cytometry using oviduct-specific marker. Chicken mutated ovalbumin promoter (Mut-4.4-kb-pOV) was validated in these cells using luciferase reporter analysis. Results: The isolated cOECs revealed that the oviduct-specific marker, ovalbumin protein, was clearly detected by immunofluorescence, western blot, and flow cytometry analysis revealed that approximately 79.40% of the cells contained this protein. Also, luciferase reporter analysis showed that the constructed Mut-4.4-kb-pOV exhibited 7.1-fold (p