Enforced activation of STAT5A facilitates the generation of embryonic stem-derived hematopoietic stem cells that contribute to hematopoiesis in vivo Journal Article
| Authors: | Schuringa, J. J.; Wu, K.; Morrone, G.; Moore, M. A. S. |
| Article Title: | Enforced activation of STAT5A facilitates the generation of embryonic stem-derived hematopoietic stem cells that contribute to hematopoiesis in vivo |
| Abstract: | Little is known about the molecular mechanisms that direct the transition from primitive to definitive hematopoiesis. In this study, we cocultured murine embryonic stem (ES) cells on OP9 stroma to induce hematopoietic differentiation as a model to study factors involved in the generation of adult hematopoietic stem cells (HSCs). Overexpression of the constitutively activated mutant signal transducer and activator of transcription (STAT) 5A(1*6) in ES cells facilitated the generation of cells that expressed the endothelial-hemangioblast marker Flk-1 within 5 days of coculture on OP9. The first CD41+/ CD45+/c-Kit+/Flk-1- hematopoietic cells arose in our culture conditions between days 5 and 7. Persistent activation of STAT5A greatly enhanced the generation of hematopoietic progenitors compared with controls, as determined by colony assays in methylcellulose. Moreover, whereas controls generated only a short transient wave of hematopoiesis lasting less than 3 weeks, expression of STAT5A(1*6) resulted in the generation of hematopoietic cobblestone area-forming cells (CAFCs) on OP9 that could be serially passaged onto new OP9, giving rise to second and third CAFCs that generated hematopoietic progenitors for ≥5 weeks, indicating a role for STAT5A in HSC self-renewal in vitro. Several definitive hematopoietic genes were upregulated by STAT5A (1*6), as well as Runx1/AML1, vascular endothelial growth factor, oncostatin M receptor, HoxB4, Wnt5A, Delta-like-1, and Bmi-1. Furthermore, ES-derived hematopoietic cells expressing STAT5A(1*6) contributed to myeloid-lymphoid hematopoiesis in primary and secondary nonobese diabetic-severe combined immunodeficiency recipients, although no donor-derived cells could be detected after 7 weeks in the secondary recipients. These data indicate that a persistent activation of STAT5A allows the generation of ES-derived HSCs that can, at least for an intermediate period, contribute to hematopoiesis in vivo. |
| Keywords: | signal transduction; vasculotropin; controlled study; dna-binding proteins; proto-oncogene proteins; nonhuman; flow cytometry; polymerase chain reaction; animal cell; mouse; phenotype; animals; mice; gene overexpression; embryo; cell line; in vivo study; cell differentiation; in vitro study; mice, scid; cell assay; vasculotropin receptor 2; vascular endothelial growth factor receptor-2; time factors; cell lineage; animalia; transcription factors; stem cell; cell culture; oligonucleotide array sequence analysis; hematopoietic system; hematopoietic stem cells; murinae; immunoblotting; cell culture techniques; stem cells; embryonic stem cells; green fluorescent proteins; electroporation; hematopoiesis; trans-activators; hematopoietic stem cell; stat5 transcription factor; self-renewal; cell separation; cd45 antigen; cell transplantation; core binding factor alpha 2 subunit; embryo cell; coculture techniques; transcription factor runx1; stat5a protein; fibrinogen receptor; definitive hematopoiesis; milk proteins; article; stat5a; methylcellulose; oncostatin m |
| Journal Title: | Stem Cells |
| Volume: | 22 |
| Issue: | 7 |
| ISSN: | 1066-5099 |
| Publisher: | AlphaMed Press |
| Date Published: | 2004-12-01 |
| Start Page: | 1191 |
| End Page: | 1204 |
| Language: | English |
| DOI: | 10.1634/stemcells.2004-0033 |
| PROVIDER: | scopus |
| PUBMED: | 15579639 |
| DOI/URL: | |
| Notes: | Stem Cells -- Cited By (since 1996):30 -- Export Date: 16 June 2014 -- CODEN: STCEE -- Source: Scopus |
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