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

STAR Protoc. 2023 Sep 15;4(3):102495. doi: 10.1016/j.xpro.2023.102495. Epub 2023 Aug 4.

Abstract

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.

Keywords: Antibody; Cell Culture; Cell Isolation; Model Organisms; Protein Expression and Purification.

MeSH terms

  • Animals
  • Chickens*
  • Epithelial Cells
  • Female
  • Humans
  • Oviducts* / metabolism
  • Povidone*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Silicon Dioxide*

Substances

  • Percoll
  • Recombinant Proteins
  • Povidone
  • Silicon Dioxide