In vitro differentiation of mouse embryonic stem cells as a model of early hematopoietic development.
Embryonic Stem (ES) are pluripotent cells derived from the inner cell mass of blastocysts. ES cells differentiate in vitro into all kind of cells and the development of endothelial and hematopoietic cells from mouse ES cells has been especially established. As such, the in vitro differentiation of ES cells provides a powerful experimental model to study and determine the role of specific genes in the development of the hematopoietic system. Using this approach we have demonstrated the critical function of the transcription factor AML1/Runx1 at the onset of hematopoietic development (Blood 100:458-466, 2002; Blood 103:886-889, 2004). In this chapter, we will describe our protocols and methods for the culture of healthy ES cells, their effective differentiation toward hematopoiesis, and the quantitative analysis of their hematopoietic potential by replating or gene expression analyses.
Sroczynska
Patrycja
P
Stem Cell Biology and Research, Paterson Institute for Cancer Research, University of Manchester, Manchester, UK.
Lancrin
Christophe
C
Pearson
Stella
S
Kouskoff
Valerie
V
Lacaud
Georges
G
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D015870:Gene Expression; D006410:Hematopoiesis / Q000502:physiology*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D016133:Polymerase Chain Reaction / Q000379:methods
D050676:Core Binding Factor Alpha 2 Subunit; D017931:DNA Primers; C493728:RUNX1 protein, human