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

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 s...

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Detalles Bibliográficos
Autores principales: Loubser, Chiron, Nikitina, Natalya V.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10432238/
https://www.ncbi.nlm.nih.gov/pubmed/37542716
http://dx.doi.org/10.1016/j.xpro.2023.102495
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author Loubser, Chiron
Nikitina, Natalya V.
author_facet Loubser, Chiron
Nikitina, Natalya V.
author_sort Loubser, Chiron
collection PubMed
description 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.
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spelling pubmed-104322382023-08-18 Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins Loubser, Chiron Nikitina, Natalya V. STAR Protoc Protocol 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. Elsevier 2023-08-04 /pmc/articles/PMC10432238/ /pubmed/37542716 http://dx.doi.org/10.1016/j.xpro.2023.102495 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Loubser, Chiron
Nikitina, Natalya V.
Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins
title Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins
title_full Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins
title_fullStr Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins
title_full_unstemmed Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins
title_short Protocol to establish an oviduct epithelial cell line derived from Gallus gallus using Percoll for in vitro validation of recombinant proteins
title_sort protocol to establish an oviduct epithelial cell line derived from gallus gallus using percoll for in vitro validation of recombinant proteins
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10432238/
https://www.ncbi.nlm.nih.gov/pubmed/37542716
http://dx.doi.org/10.1016/j.xpro.2023.102495
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