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Blood, Vol. 95 No. 3 (February 1), 2000:
pp. 855-862
High-resolution tracking of cell division suggests similar cell
cycle kinetics of hematopoietic stem cells stimulated in vitro and
in vivo
Robert A. J. Oostendorp,
Julie Audet, and
Connie
J. Eaves
From the Terry Fox Laboratory, British Columbia Cancer Agency,
Vancouver, BC, Canada; the Department of Medical Genetics and the
Biotechnology Laboratory, University of British Columbia, Vancouver,
BC, Canada; and the Department of Cell Biology and Genetics, Erasmus
University, Rotterdam, the Netherlands.
The kinetics of proliferation of primitive murine bone marrow (BM)
cells stimulated either in vitro with growth factors (fetal liver
tyrosine kinase ligand 3 [FL], Steel factor [SF], and
interleukin-11 [IL-11], or hyper-IL-6) or in vivo by factors active
in myeloablated recipients were examined. Cells were first labeled with
5- and 6-carboxyfluorescein diacetate succinimidyl ester (CFSE) and
then incubated overnight prior to isolating CFSE+ cells.
After 2 more days in culture, more than 90% of the in vivo
lymphomyeloid repopulating activity was associated with the most
fluorescent CFSE+ cells (ie, cells that had not yet
divided), although this accounted for only 25% of the repopulating
stem cells measured in the CFSE+ "start"
population. After a total of 4 days in culture (1 day later), 15-fold
more stem cells were detected (ie, 4-fold more than the day 1 input
number), and these had become (and thereafter remained) exclusively
associated with cells that had divided at least once in vitro. Flow
cytometric analysis of CFSE+ cells recovered from the BM
of transplanted mice indicated that these cells proliferated slightly
faster (up to 5 divisions completed within 2 days and up to 8 divisions
completed within 3 days in vivo versus 5 and 7 divisions, respectively,
in vitro). FL, SF, and ligands which activate gp130 are thus efficient
stimulators of transplantable stem cell self-renewal divisions in
vitro. The accompanying failure of these cells to accumulate rapidly
indicates important changes in their engraftment potential independent
of accompanying changes in their differentiation status.
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