Lentiviral transduction and clonal selection of hESCs with endothelial-specific transgenic reporters. Academic Article uri icon

Overview

MeSH

  • Cell Line
  • Humans
  • Organ Specificity

MeSH Major

  • Embryonic Stem Cells
  • Endothelial Cells
  • Genes, Reporter
  • Lentivirus
  • Transduction, Genetic
  • Transgenes

abstract

  • Generation of vascular endothelial cells (EC) from human embryonic stem cells (hESC) is a vital component of cell-based strategies for treatment of cardiovascular disease. Before hESC-derived ECs can be administered in therapeutic modalities, however, chemically defined culture conditions must be developed that reproducibly and robustly induce vascular differentiation. One approach to screening for culture conditions that support differentiation of hESCs to any cell type is their genetic modification with exogenous DNA sequence comprising a tissue-specific gene promoter driving reporters such as fluorescent protein or antibiotic drug resistance. The protocols herein provide instructions for the generation of clonal hESC lines containing a reporter transgene that is specifically expressed in vascular endothelial derivatives. Additionally, they demonstrate the methodology employed to assess vascular differentiation from clonal lines. Together, these protocols provide a solid foundation for study of vascular differentiation, and may also be applied, in principle, to studies of other specialized cell types derived from hESCs. Copyright © 2011 by John Wiley & Sons, Inc.

publication date

  • April 2011

has subject area

  • Cell Line
  • Embryonic Stem Cells
  • Endothelial Cells
  • Genes, Reporter
  • Humans
  • Lentivirus
  • Organ Specificity
  • Transduction, Genetic
  • Transgenes

Research

keywords

  • Journal Article

Identity

Language

  • eng

Digital Object Identifier (DOI)

  • 10.1002/9780470151808.sc01f12s17

PubMed ID

  • 21509935

Additional Document Info

start page

  • Unit1F.12

volume

  • Chapter 1