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Prepublished online as a Blood First Edition Paper on July 12, 2002; DOI 10.1182/blood-2002-05-1511.
Submitted May 29, 2002
Accepted June 12, 2002
Identification of the hemangioblast in post-natal life
Elvira Pelosi, Mauro Valtieri, Simona Coppola, Rosanna Botta, Marco Gabbianelli, Valentina Lulli, Giovanna Marziali, Barbara Masella, Robert Muller, Cecilia Sgadari, Ugo Testa, Giuseppina Bonanno, and Cesare Peschle*
Hematology-Oncology, Istituto Superiore di Sanita', Rome, Italy, Italy
Hematology-Oncology, Istituto Superiore di Sanita', Rome, Italy, Italy; Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA
Kimmel Cancer Center, Thomas Jefferson University, Philadelphia, PA, USA
Virology, Istituto Superiore di Sanita', Rome, Italy, Italy
Obstetrics and Gynaecology, Catholic University, Rome, Italy, Italy
* Corresponding author; email: cesare.peschle{at}mail.tju.edu.
Post-natal CD34+ cells expressing KDR (vascular endothelial growth factor receptor 2) generate hematopoietic or endothelial progeny in different in vitro/in vivo assays. Hypothetically, CD34+KDR+ cells may comprise hemangioblasts bipotent for both lineages. This hypothesis is consistent with two series of experiments. (i) In clonogenic culture permissive for both hematopoietic and endothelial cell growth, CD34+KDR+ cells generate large hemato-endothelial (Hem-End) colonies (5% of seeded cells), whereas CD34+KDR- cells do not. Limiting dilution analysis indicates that Hem-End colonies are clonally generated by single hemangioblasts. Sibling cells generated by a hemangioblast, replated in unicellular culture, produce either hematopoietic or Hem-End colonies, depending on the specific culture conditions. Identification of endothelial cell was based on expression of VE-cadherin and endothelial markers, together with lack of CD45 and hematopoietic molecules, as evaluated by immunofluorescence, immunocytochemistry and RT-PCR; furthermore, endothelial cell were functionally identified by low density lipoprotein (LDL) uptake and tube formation assays. (ii) To evaluate the self-renewal capacity of hemangioblasts, single CD34+KDR+ cells were grown in 3-month-extended long-term culture (ELTC) through three serial culture rounds, i.e., blast cells generated in unicellular ELTC were reseeded for a subsequent round of unicellular ELTC. After 9 months, 10% blasts from tertiary ELTC functioned as hemangioblasts and generated macroscopic Hem-End colonies in clonogenic culture. These studies identify post-natal hemangioblasts in a CD34+KDR+ cell subset, endowed with long term proliferative potential and bilineage differentiation capacity. Although exceedingly rare, hemangioblasts may represent the lifetime source/reservoir for primitive hematopoietic and endothelial progenitors.
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