A human VE-cadherin-tdTomato and CD43-green fluorescent protein dual reporter cell line for study endothelial to hematopoietic transition

Stem Cell Res. 2016 Sep;17(2):401-405. doi: 10.1016/j.scr.2016.09.004. Epub 2016 Sep 13.

Abstract

Human embryonic stem cell line WA01 was genetically modified using zinc-finger nucleases and the PiggyBac/transponson system to introduce a fluorescence reporter for VE-cadherin (VEC; tdTomato) and CD43 (eGFP). Phenotypic and functional assays for pluripotency revealed the modified hES cell reporter lines remained normal. When the cells were differentiated into hematoendothelial lineages, either by directed differentiation or direct reprogramming, flow cytometric and fluorescence microscopy showed that VEC+ endothelial cells express tdTomato and CD43+ hematopoietic progenitors express eGFP.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, CD / genetics
  • Antigens, CD / metabolism*
  • Cadherins / genetics
  • Cadherins / metabolism*
  • Cell Differentiation
  • Cells, Cultured
  • Embryoid Bodies / metabolism
  • Embryoid Bodies / pathology
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism
  • Genes, Reporter
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • Hematopoietic Stem Cells / cytology*
  • Hematopoietic Stem Cells / metabolism
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / metabolism
  • Humans
  • Karyotype
  • Leukosialin / genetics
  • Leukosialin / metabolism*
  • Male
  • Microscopy, Fluorescence
  • Time-Lapse Imaging
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Antigens, CD
  • Cadherins
  • Leukosialin
  • Transcription Factors
  • cadherin 5
  • Green Fluorescent Proteins