GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering

Abstract Engineered epithelial cell sheets for clinical replacement of non-functional upper aerodigestive tract mucosa are regulated as medicinal products and should be manufactured to the standards of good manufacturing practice (GMP). The current gold standard for growth of epithelial cells for re...

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Autores principales:	Angela Tait, Toby Proctor, Nick J. I. Hamilton, Martin A. Birchall, Mark W. Lowdell
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8f2673ced80b44e8beb594861f87e6b8
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spelling	oai:doaj.org-article:8f2673ced80b44e8beb594861f87e6b82021-12-02T17:52:31ZGMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering10.1038/s41598-021-91939-02045-2322https://doaj.org/article/8f2673ced80b44e8beb594861f87e6b82021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91939-0https://doaj.org/toc/2045-2322Abstract Engineered epithelial cell sheets for clinical replacement of non-functional upper aerodigestive tract mucosa are regulated as medicinal products and should be manufactured to the standards of good manufacturing practice (GMP). The current gold standard for growth of epithelial cells for research utilises growth arrested murine 3T3 J2 feeder layers, which are not available for use as a GMP compliant raw material. Using porcine mucosal tissue, we demonstrate a new method for obtaining and growing non-keratinised squamous epithelial cells and fibroblast cells from a single biopsy, replacing the 3T3 J2 with a growth arrested primary fibroblast feeder layer and using pooled Human Platelet lysate (HPL) as the media serum supplement to replace foetal bovine serum (FBS). The initial isolation of the cells was semi-automated using an Octodissociator and the resultant cell suspension cryopreservation for future use. When compared to the gold standard of 3T3 J2 and FBS containing medium there was no reduction in growth, viability, stem cell population or ability to differentiate to mature epithelial cells. Furthermore, this method was replicated with Human buccal tissue, providing cells of sufficient quality and number to create a tissue engineered sheet.Angela TaitToby ProctorNick J. I. HamiltonMartin A. BirchallMark W. LowdellNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Angela Tait Toby Proctor Nick J. I. Hamilton Martin A. Birchall Mark W. Lowdell GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering
description	Abstract Engineered epithelial cell sheets for clinical replacement of non-functional upper aerodigestive tract mucosa are regulated as medicinal products and should be manufactured to the standards of good manufacturing practice (GMP). The current gold standard for growth of epithelial cells for research utilises growth arrested murine 3T3 J2 feeder layers, which are not available for use as a GMP compliant raw material. Using porcine mucosal tissue, we demonstrate a new method for obtaining and growing non-keratinised squamous epithelial cells and fibroblast cells from a single biopsy, replacing the 3T3 J2 with a growth arrested primary fibroblast feeder layer and using pooled Human Platelet lysate (HPL) as the media serum supplement to replace foetal bovine serum (FBS). The initial isolation of the cells was semi-automated using an Octodissociator and the resultant cell suspension cryopreservation for future use. When compared to the gold standard of 3T3 J2 and FBS containing medium there was no reduction in growth, viability, stem cell population or ability to differentiate to mature epithelial cells. Furthermore, this method was replicated with Human buccal tissue, providing cells of sufficient quality and number to create a tissue engineered sheet.
format	article
author	Angela Tait Toby Proctor Nick J. I. Hamilton Martin A. Birchall Mark W. Lowdell
author_facet	Angela Tait Toby Proctor Nick J. I. Hamilton Martin A. Birchall Mark W. Lowdell
author_sort	Angela Tait
title	GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering
title_short	GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering
title_full	GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering
title_fullStr	GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering
title_full_unstemmed	GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering
title_sort	gmp compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/8f2673ced80b44e8beb594861f87e6b8
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_version_	1718379178911334400

GMP compliant isolation of mucosal epithelial cells and fibroblasts from biopsy samples for clinical tissue engineering

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