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Prepublished online as a Blood First Edition Paper on July 31, 2003; DOI 10.1182/blood-2003-03-0729.
Submitted March 7, 2003
Accepted July 21, 2003
Primary endothelial cells isolated from the yolk sac and para-aortic splanchnopleura (P-Sp) support the expansion of adult marrow stem cells in vitro
Weiming Li, Scott A Johnson, William C Shelley, Michael Ferkowicz, Paul Morrison, Yanjun Li, and Mervin C Yoder*
Department of Pediatrics, Herman B Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN, USA
* Corresponding author; email: myoder{at}iupui.edu.
The embryonic origin and development of hematopoietic and endothelial cells is highly interdependent. We hypothesized that primary endothelial cells from murine yolk sac and para-aortic splanchnopleura (P-Sp) may possess the capacity to expand hematopoietic stem (HSC) and progenitor cells ex vivo. Using Tie2-GFP transgenic mice in combination with fluorochrome-conjugated monoclonal antibodies to vascular endothelial growth factor receptor-2 (Flk-1) and CD41, we have successfully isolated pure populations of primary endothelial cells from 9.5 days post coitum (dpc) yolk sac and P-Sp. Adult murine bone marrow Sca-1+c-Kit+lin- cells were co-cultured with yolk sac or P-Sp Tie2-GFP+Flk-1+CD41- endothelial cell monolayers for seven days and the total number of nonadherent cells increased 47 and 295 fold, respectively, and hematopoietic progenitor counts increased 9.4 and 11.4 fold, respectively. Both the yolk sac and P-Sp endothelial cell co-cultures facilitated long-term (>6 months) HSC competitive repopulating ability (2.8 to 9.8 fold increases, respectively). These data suggest that 9.5 dpc yolk sac and P-Sp-derived primary Tie2-GFP+Flk-1+CD41- endothelial cells possess the capacity to expand adult bone marrow hematopoietic progenitor cells and HSC repopulating ability ex vivo.
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