Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells

Abstract Bovine embryonic stem cells (bESCs) extend the lifespan of the transient pluripotent bovine inner cell mass in vitro. After years of research, derivation of stable bESCs was only recently reported. Although successful, bESC culture relies on complex culture conditions that require a custom-...

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Autores principales:	Delia Alba Soto, Micaela Navarro, Canbin Zheng, Michelle Margaret Halstead, Chuan Zhou, Carly Guiltinan, Jun Wu, Pablo Juan Ross
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/227ee01bdda346c98bb2a5124deb5b52
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spelling	oai:doaj.org-article:227ee01bdda346c98bb2a5124deb5b522021-12-02T16:53:19ZSimplification of culture conditions and feeder-free expansion of bovine embryonic stem cells10.1038/s41598-021-90422-02045-2322https://doaj.org/article/227ee01bdda346c98bb2a5124deb5b522021-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-90422-0https://doaj.org/toc/2045-2322Abstract Bovine embryonic stem cells (bESCs) extend the lifespan of the transient pluripotent bovine inner cell mass in vitro. After years of research, derivation of stable bESCs was only recently reported. Although successful, bESC culture relies on complex culture conditions that require a custom-made base medium and mouse embryonic fibroblasts (MEF) feeders, limiting the widespread use of bESCs. We report here simplified bESC culture conditions based on replacing custom base medium with a commercially available alternative and eliminating the need for MEF feeders by using a chemically-defined substrate. bESC lines were cultured and derived using a base medium consisting of N2B27 supplements and 1% BSA (NBFR-bESCs). Newly derived bESC lines were easy to establish, simple to propagate and stable after long-term culture. These cells expressed pluripotency markers and actively proliferated for more than 35 passages while maintaining normal karyotype and the ability to differentiate into derivatives of all three germ lineages in embryoid bodies and teratomas. In addition, NBFR-bESCs grew for multiple passages in a feeder-free culture system based on vitronectin and Activin A medium supplementation while maintaining pluripotency. Simplified conditions will facilitate the use of bESCs for gene editing applications and pluripotency and lineage commitment studies.Delia Alba SotoMicaela NavarroCanbin ZhengMichelle Margaret HalsteadChuan ZhouCarly GuiltinanJun WuPablo Juan RossNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Delia Alba Soto Micaela Navarro Canbin Zheng Michelle Margaret Halstead Chuan Zhou Carly Guiltinan Jun Wu Pablo Juan Ross Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells
description	Abstract Bovine embryonic stem cells (bESCs) extend the lifespan of the transient pluripotent bovine inner cell mass in vitro. After years of research, derivation of stable bESCs was only recently reported. Although successful, bESC culture relies on complex culture conditions that require a custom-made base medium and mouse embryonic fibroblasts (MEF) feeders, limiting the widespread use of bESCs. We report here simplified bESC culture conditions based on replacing custom base medium with a commercially available alternative and eliminating the need for MEF feeders by using a chemically-defined substrate. bESC lines were cultured and derived using a base medium consisting of N2B27 supplements and 1% BSA (NBFR-bESCs). Newly derived bESC lines were easy to establish, simple to propagate and stable after long-term culture. These cells expressed pluripotency markers and actively proliferated for more than 35 passages while maintaining normal karyotype and the ability to differentiate into derivatives of all three germ lineages in embryoid bodies and teratomas. In addition, NBFR-bESCs grew for multiple passages in a feeder-free culture system based on vitronectin and Activin A medium supplementation while maintaining pluripotency. Simplified conditions will facilitate the use of bESCs for gene editing applications and pluripotency and lineage commitment studies.
format	article
author	Delia Alba Soto Micaela Navarro Canbin Zheng Michelle Margaret Halstead Chuan Zhou Carly Guiltinan Jun Wu Pablo Juan Ross
author_facet	Delia Alba Soto Micaela Navarro Canbin Zheng Michelle Margaret Halstead Chuan Zhou Carly Guiltinan Jun Wu Pablo Juan Ross
author_sort	Delia Alba Soto
title	Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells
title_short	Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells
title_full	Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells
title_fullStr	Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells
title_full_unstemmed	Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells
title_sort	simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/227ee01bdda346c98bb2a5124deb5b52
work_keys_str_mv	AT deliaalbasoto simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells AT micaelanavarro simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells AT canbinzheng simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells AT michellemargarethalstead simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells AT chuanzhou simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells AT carlyguiltinan simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells AT junwu simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells AT pablojuanross simplificationofcultureconditionsandfeederfreeexpansionofbovineembryonicstemcells
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Simplification of culture conditions and feeder-free expansion of bovine embryonic stem cells

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