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Blood, Vol. 92 No. 8 (October 15), 1998:
pp. 2641-2649
RAPID COMMUNICATION
Cell Cycle-Related Changes in Repopulating Capacity of Human Mobilized
Peripheral Blood CD34+ Cells in Non-Obese Diabetic/Severe
Combined Immune-Deficient Mice
André Gothot,
Johannes C.M. van der Loo,
D. Wade Clapp, and
Edward F. Srour
From the Division of Hematology/Oncology and Indiana Elks Cancer
Research Center, Department of Medicine, and Department of Pediatrics,
Herman B Wells Center for Pediatric Research, Indiana School of
Medicine, Indianapolis, IN.
Most primitive hematopoietic progenitor cells reside in vivo within
the G0/G1 phase of the cell cycle. By
simultaneous DNA/RNA staining it is possible to distinguish
G0 and G1 states and to isolate cells in
defined phases of the cell cycle. We report here the use of cell cycle
fractionation to separate human mobilized peripheral blood (MPB)
CD34+ cells capable of repopulating the bone marrow (BM)
of non-obese diabetic/severe combined immune-deficient (NOD/SCID) mice.
In freshly isolated MPB, repopulating cells were predominant within the
G0 phase, because transplantation of CD34+
cells residing in G0 (G0CD34+)
resulted on average in a 16.6- ± 3.2-fold higher BM chimerism than
infusion of equal numbers of CD34+ cells isolated in
G1. We then investigated the effect of ex vivo cell cycle
progression, in the absence of cell division, on engraftment capacity.
Freshly isolated G0CD34+ cells were activated
by interleukin-3 (IL-3), stem cell factor (SCF), and flt3-ligand (FL)
for a 36-hour incubation period during which a fraction of cells
progressed from G0 into G1 but did not complete
a cell cycle. The repopulating capacity of stimulated cells was
markedly diminished compared with that of unmanipulated G0CD34+ cells. Cells that remained in
G0 during the 36-hour incubation period and those that
traversed into G1 were sorted and assayed separately in
NOD/SCID recipients. The repopulating ability of cells remaining in
G0 was insignificantly reduced compared with that of
unstimulated G0CD34+ cells. On the contrary,
CD34+ cells traversing from G0 into
G1 were largely depleted of repopulating capacity. Similar
results were obtained when G0CD34+ cells were
activated by the combination of thrombopoietin-SCF-FL. These studies
provide direct evidence of the quiescent nature of cells capable of
repopulating the BM of NOD/SCID mice. Furthermore, these data also
demonstrate that G0-G1 progression in vitro is associated with a decrease in engraftment capacity.
© 1998 by The American Society of Hematology.
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