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Prepublished online as a Blood First Edition Paper on June 28, 2002; DOI 10.1182/blood-2002-02-0490.
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Blood, 1 December 2002, Vol. 100, No. 12, pp. 3975-3982
HEMATOPOIESIS
The spleen is a major site of megakaryopoiesis following
transplantation of murine hematopoietic stem cells
William B. Slayton,
Ann Georgelas,
L. Jeanne Pierce,
Kojo S. Elenitoba-Johnson,
S. Scott Perry,
Melissa Marx, and
Gerald J. Spangrude
From the Departments of Pediatrics, Pathology, and
Oncological Sciences, University of Utah School of Medicine, Salt Lake
City.
The stem cell pool can be fractionated by using the mitochondrial
dye, rhodamine-123, into Rholow hematopoietic stem cells
and Rhohigh progenitors. Rholow stem cells
permanently engraft all lineages, whereas Rhohigh
progenitors transiently produce erythrocytes, without
substantial platelet or granulocyte production. We hypothesized
that the inability of the Rhohigh cells to produce
platelets in vivo was due to the fact that these cells preferentially
engraft in the spleen and lack marrow engraftment. Initially, we
demonstrated that Rhohigh progenitors produced more
megakaryocytes in vitro than Rholow stem cells did. To
study the activity of the Rholow and Rhohigh
subsets in vivo, we used mice allelic at the hemoglobin and glucose phosphate isomerase loci to track donor-derived erythropoiesis and
thrombopoiesis. Rholow stem cells contributed to robust and
long-term erythroid and platelet engraftment, whereas
Rhohigh progenitors contributed only to transient erythroid
engraftment and produced very low numbers of platelets in vivo.
Donor-derived megakaryopoiesis occurred at higher densities in the
spleen than in the bone marrow in animals receiving Rholow
stem cells and peaked around day 28. Blockade of splenic engraftment using pertussis toxin did not affect the peak of splenic
megakaryopoiesis, supporting the hypothesis that these megakaryocytes
were derived from progenitors that originated in the bone marrow. These
data emphasize that in vitro behavior of hematopoietic progenitor cell subsets does not always predict their behavior following
transplantation. This study supports a major role for the spleen in
thrombopoiesis following engraftment of transplanted stem cells in
irradiated mice.
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