Hematopoietic-specific genes are not induced during in vitro differentiation of scl-null embryonic stem cells

Andrew G. Elefanty, Lorraine Robb, Raquella Birner, C. Glenn Begley

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The helix-loop-helix transcription factor, scl, plays an essential role in hematopoletic development. Embryos in which the gene has been disrupted fail to develop yolk sac erythropoiesis, and sc/-null embryonic stem cells do not contribute to hematopoiesis in chimeric mice. To analyze the molecular consequences of scl deficiency, we compared the gene expression profiles of control (wild-type and scl-heterozygous) and scl-null embryonic stem cells differentiated in vitro for up to 12 days. In control and scl-null embryoid bodies the temporal expression pattern of genes associated with the formation of ventral mesoderm, such as Brachyury, bone morphogenetic protein-4, and flk-1, was identical. Similarly, GATA-2, CD34, and c-kit, which are coexpressed in endothelial and hematopoietic lineages, were expressed normally inscl-null embryonic stem cell lines. However, hemetopoieticrestricted genes, including the transcription factors GATA-1, EKLF, and PU.1 as well as globin genes and myeloperoxidase, were only expressed in wild-type and scl-heterozygous embryonic stem cells. Indirect immunofluorescence was used to confirm the observations that GATA-1 and globins were only present in control embryoid bodies but that CD34 was found on both control and scl-null embryoid bodies. These data extend the previous gene ablation studies and support a model whereby scl is absolutely required for commitment of a putative hemangioblast to the hematopoietic lineage but that it is dispensable for endothelial differentiation.

Original languageEnglish
Pages (from-to)1435-1447
Number of pages13
Issue number4
Publication statusPublished - 15 Aug 1997
Externally publishedYes

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