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Prepublished online as a Blood First Edition Paper on July 12, 2002; DOI 10.1182/blood-2002-05-1511.
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Blood, 1 November 2002, Vol. 100, No. 9, pp. 3203-3208
HEMATOPOIESIS
Identification of the hemangioblast in postnatal life
Elvira Pelosi,
Mauro Valtieri,
Simona Coppola,
Rosanna Botta,
Marco Gabbianelli,
Valentina Lulli,
Giovanna Marziali,
Barbara Masella,
Robert Müller,
Cecilia Sgadari,
Ugo Testa,
Giuseppina Bonanno, and
Cesare Peschle
From the Kimmel Cancer Center, Thomas Jefferson
University, Philadelphia, PA; the Departments of Hematology-Oncology
and Virology, Istituto Superiore di Sanità, Rome; and the
Department of Obstetrics and Gynecology, Catholic University, Rome,
Italy.
Postnatal CD34+ cells expressing vascular
endothelial growth factor receptor 2 (KDR) generate hematopoietic or
endothelial progeny in different in vitro and in vivo assays.
Hypothetically, CD34+KDR+ cells may comprise
hemangioblasts bipotent for both lineages. This hypothesis is
consistent with 2 series of experiments. In the first series, in
clonogenic culture permissive for 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 cells was based on the expression of
VE-cadherin and endothelial markers and with lack of CD45 and
hematopoietic molecules, as evaluated by immunofluorescence,
immunocytochemistry, and reverse transcription-polymerase chain
reaction. Furthermore, endothelial cells were functionally identified
using low-density lipoprotein (LDL) uptake and tube-formation assays.
In the second series, to evaluate the self-renewal capacity of
hemangioblasts, single CD34+KDR+ cells were
grown in 3-month extended long-term culture (ELTC) through 3 serial
culture rounds that is, 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 identified postnatal 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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