Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins

• Published in: STAR protocols 2023-09, Vol.4 (3), p.102495-102495, Article 102495
• Main Authors: Loubser, Chiron, Nikitina, Natalya V.
• Format: Article
• Language: English
• Subjects: Animals; Antibody; Cell Culture; Cell Isolation; Chickens; Epithelial Cells; Female; Humans; Model Organisms; Oviducts - metabolism; Povidone; Protein Expression and Purification; Protocol; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Silicon Dioxide
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2023.102495

In vitro validation of therapeutic and recombinant proteins expressed from transgenic chickens is limited by the co-culture of fibroblasts. Here, we present a protocol for isolating pure epithelial cells derived from the magnum tubular glands of the chicken oviduct. We describe steps for preparing solutions and buffers, tissue collection, processing, dissociation, and Percoll density centrifugation to separate the epithelial cells from co-isolated fibroblasts. We then detail procedures for expressing a recombinant IgG antibody in the Percoll-derived epithelial cell line. [Display omitted] •Establishment of the chicken oviduct cell line derived from the magnum•Separation of co-isolated fibroblasts and epithelial cells with Percoll•Immunostaining for OVA as confirmation of cell identity•PEI Max transfection, expression, and purification of recombinant IgG antibody Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. In vitro validation of therapeutic and recombinant proteins expressed from transgenic chickens is limited by the co-culture of fibroblasts. Here, we present a protocol for isolating pure epithelial cells derived from the magnum tubular glands of the chicken oviduct. We describe steps for preparing solutions and buffers, tissue collection, processing, dissociation, and Percoll density centrifugation to separate the epithelial cells from co-isolated fibroblasts. We then detail procedures for expressing a recombinant IgG antibody in the Percoll-derived epithelial cell line.