Cargando…

PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines

Extraembryonic endoderm stem (XEN) cell lines can be derived and maintained in vitro and reflect the primitive endoderm lineage. Platelet-derived growth factor receptor alpha (PDGFRA) is thought to be essential for the derivation and maintenance of mouse XEN cell lines. Here, we have re-evaluated th...

Descripción completa

Detalles Bibliográficos
Autores principales: Lin, Jiangwei, Khan, Mona, Zapiec, Bolek, Mombaerts, Peter
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5639183/
https://www.ncbi.nlm.nih.gov/pubmed/28919262
http://dx.doi.org/10.1016/j.stemcr.2017.08.005
_version_ 1783270844560048128
author Lin, Jiangwei
Khan, Mona
Zapiec, Bolek
Mombaerts, Peter
author_facet Lin, Jiangwei
Khan, Mona
Zapiec, Bolek
Mombaerts, Peter
author_sort Lin, Jiangwei
collection PubMed
description Extraembryonic endoderm stem (XEN) cell lines can be derived and maintained in vitro and reflect the primitive endoderm lineage. Platelet-derived growth factor receptor alpha (PDGFRA) is thought to be essential for the derivation and maintenance of mouse XEN cell lines. Here, we have re-evaluated this requirement for PDGFRA. We derived multiple PDGFRA-deficient XEN cell lines from postimplantation and preimplantation embryos of a PDGFRA-GFP knockout strain. We also converted PDGFRA-deficient embryonic stem cell lines into XEN cell lines chemically by transient culturing with retinoic acid and Activin A. We confirmed the XEN profile of our 12 PDGFRA-deficient cell lines by immunofluorescence with various markers, by NanoString gene expression analyses, and by their contribution to the extraembryonic endoderm of chimeric embryos produced by injecting these cells into blastocysts. Thus, PDGFRA is not essential for the derivation and maintenance of XEN cell lines.
format Online
Article
Text
id pubmed-5639183
institution National Center for Biotechnology Information
language English
publishDate 2017
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-56391832017-10-20 PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines Lin, Jiangwei Khan, Mona Zapiec, Bolek Mombaerts, Peter Stem Cell Reports Report Extraembryonic endoderm stem (XEN) cell lines can be derived and maintained in vitro and reflect the primitive endoderm lineage. Platelet-derived growth factor receptor alpha (PDGFRA) is thought to be essential for the derivation and maintenance of mouse XEN cell lines. Here, we have re-evaluated this requirement for PDGFRA. We derived multiple PDGFRA-deficient XEN cell lines from postimplantation and preimplantation embryos of a PDGFRA-GFP knockout strain. We also converted PDGFRA-deficient embryonic stem cell lines into XEN cell lines chemically by transient culturing with retinoic acid and Activin A. We confirmed the XEN profile of our 12 PDGFRA-deficient cell lines by immunofluorescence with various markers, by NanoString gene expression analyses, and by their contribution to the extraembryonic endoderm of chimeric embryos produced by injecting these cells into blastocysts. Thus, PDGFRA is not essential for the derivation and maintenance of XEN cell lines. Elsevier 2017-09-14 /pmc/articles/PMC5639183/ /pubmed/28919262 http://dx.doi.org/10.1016/j.stemcr.2017.08.005 Text en © 2017 The Author(s) http://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 Report
Lin, Jiangwei
Khan, Mona
Zapiec, Bolek
Mombaerts, Peter
PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines
title PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines
title_full PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines
title_fullStr PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines
title_full_unstemmed PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines
title_short PDGFRA Is Not Essential for the Derivation and Maintenance of Mouse Extraembryonic Endoderm Stem Cell Lines
title_sort pdgfra is not essential for the derivation and maintenance of mouse extraembryonic endoderm stem cell lines
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5639183/
https://www.ncbi.nlm.nih.gov/pubmed/28919262
http://dx.doi.org/10.1016/j.stemcr.2017.08.005
work_keys_str_mv AT linjiangwei pdgfraisnotessentialforthederivationandmaintenanceofmouseextraembryonicendodermstemcelllines
AT khanmona pdgfraisnotessentialforthederivationandmaintenanceofmouseextraembryonicendodermstemcelllines
AT zapiecbolek pdgfraisnotessentialforthederivationandmaintenanceofmouseextraembryonicendodermstemcelllines
AT mombaertspeter pdgfraisnotessentialforthederivationandmaintenanceofmouseextraembryonicendodermstemcelllines