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Next Article 
Blood, Vol. 94 No. 7 (October 1), 1999:
pp. 2161-2168
Direct Evidence for Multiple Self-Renewal Divisions of Human In Vivo
Repopulating Hematopoietic Cells in Short-Term Culture
H. Glimm and
C.J. Eaves
From the Terry Fox Laboratory, British Columbia Cancer Agency; and
the Department of Medical Genetics, University of British Columbia,
Vancouver, BC, Canada.
Recently, culture conditions that stimulate the proliferation of
primitive hematopoietic cells defined by various phenotypic and
functional endpoints in vitro have been identified. However, evidence
that they support a high probability of self-renewal leading to a large
net expansion in vitro of transplantable cells with lympho-myeloid
repopulating ability has been more difficult to obtain. The present
study was designed to investigate whether the low overall expansion of
human repopulating hematopoietic cells seen in vitro reflects a
selective unresponsiveness of these rare cells to the growth factors
currently used to stimulate them or, alternatively, whether they do
proliferate in vitro but lose engrafting potential. For this, we used a
high-resolution procedure for tracking and reisolating cells as a
function of their proliferation history based on the loss of
cellular fluorescence after staining with (5- and 6-)
carboxyfluorescein diacetate succinimidyl ester. The results show that
the vast majority of long-term culture-initiating cells and in vivo
lympho-myeloid competitive repopulating units present in 5-day
suspension cultures initiated with CD34+ human cord blood
and fetal liver cells are the progeny of cells that have divided at
least once in response to stimulation by interleukin-3, interleukin-6,
granulocyte colony-stimulating factor, Steel factor, and Flt3-ligand.
Thus, most human repopulating cells from these two sources are
stimulated to undergo multiple divisions under currently used
short-term suspension culture conditions and a proportion of these
retain engraftment potential.
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Related Letter in Blood Online:
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Proliferative history and hematopoietic function of ex vivo expanded human CD34+ cells
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Blood 2000 96: 1609-1612.
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