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Prepublished online as a Blood First Edition Paper on September 26, 2002; DOI 10.1182/blood-2002-05-1468.
Submitted May 20, 2002
Accepted September 18, 2002
Thrombopoietin promotes mixed lineage and megakarocytic colony forming cell growth but inhibits primitive and definitive erythropoiesis in cells isolated from early murine yolk sacs
Xiaodong Xie, Rebecca J Chan, Scott A Johnson, Mark Starr, Jennifer McCarthy, Reuben Kapur, and Mervin C Yoder*
Department of Pediatrics, Indiana University, Indianapolis, IN, USA
* Corresponding author; email: myoder{at}iupui.edu.
The role of thrombopoietin (Tpo) in promoting hematopoiesis has been extensively studied in late fetal, neonatal, and adult mice. However, the effects of Tpo on early yolk sac hematopoiesis have been largely unexplored. We examined whole embryos or the cells isolated from embryo proper and yolk sacs and identified both Tpo and c-mpl (Tpo receptor) mRNA transcripts in tissues as early as embryonic day 6.5 (E6.5). Pre-somite whole embryos and somite staged yolk sac and embryo proper cells were plated in methycellulose cultures and treated with selected hematopoietic growth factors in the presence or absence of Tpo. Tpo alone failed to promote CFU formation. However, in the presence of other growth factors, Tpo caused a significant dose dependent reduction in primitive and definitive erythroid CFU growth in cultures containing E7.5 and E8.0 whole embryos and E8.25-9.5 yolk sac-derived cells. Meanwhile, Tpo treatment resulted in a significant dose dependent increase in CFU-mixed lineage (CFU-Mix) and CFU-megakaryocyte (CFU-Meg) formation in cultures containing cells from similar staged tissues. Addition of Tpo to cultures of sorted E9.5 yolk sac c-Kit+CD34+ hematopoietic progenitors also inhibited erythroid CFU growth but augmented CFU-Mix and CFU-Meg colony forming activity. Tpo effects on CFU growth were blocked in the presence of a monoclonal antibody with Tpo neutralizing activity but not with control antibody. Thus, under certain growth factor conditions, Tpo directly inhibits early yolk sac erythroid CFU growth, but facilitates megakaryocyte and mixed lineage colony formation.
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