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

Stem Cell Reports. 2017 Oct 10;9(4):1062-1070. doi: 10.1016/j.stemcr.2017.08.005. Epub 2017 Sep 14.

Abstract

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.

Keywords: Pdgfra; Sox17; XEN; postimplantation; primitive endoderm.

MeSH terms

  • Animals
  • Blastocyst / cytology
  • Blastocyst / metabolism
  • Cell Differentiation / genetics*
  • Cell Line
  • Cell Self Renewal / genetics*
  • Cluster Analysis
  • Embryo, Mammalian / cytology
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism*
  • Endoderm / cytology*
  • Gene Expression
  • Gene Expression Profiling
  • Gene Knockout Techniques
  • Genes, Reporter
  • Genotype
  • Mice
  • Receptor, Platelet-Derived Growth Factor alpha / genetics*
  • Receptor, Platelet-Derived Growth Factor alpha / metabolism

Substances

  • Receptor, Platelet-Derived Growth Factor alpha