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Ex vivo expansion and maturation of peripheral blood CD34+ cells into the
myeloid lineage
DN Haylock, LB To, TL Dowse, CA Juttner and PJ Simmons
Leukaemia Research Unit, Hanson Centre for Cancer Research, Adelaide, South
Australia.
Hematopoietic reconstitution (HR) after peripheral blood stem cell
transplantation is characterized by a delay of 8 and 12 days for recovery
to safe levels of neutrophils and platelets even in patients with the most
rapid engraftment. We postulate that a further enhancement in the rate of
HR may be achieved by transplanting with an expanded postprogenitor cell
population that can provide mature functional cells within days of
infusion. In this study we investigated the ability of combinations of
hematopoietic growth factors (HGF) to generate nascent
granulocyte-macrophage colony-forming units (CFU-GM) in a 7-day suspension
culture of peripheral blood CD34+ cells. A combination of 6 HGF, ie,
interleukin-1 beta (IL-1), IL-3, IL-6, granulocyte colony-stimulating
factor (G-CSF), granulocyte-macrophage- CSF (GM-CSF), and stem cell factor
(SCF), was identified as the most potent combination of those tested.
Subsequently, large volume suspension cultures of CD34+ cells from the same
patients using the same 6-factor combination were established and monitored
for 21 days. An exponential rate of nucleated cell production (mean
1,324-fold increase) occurred during culture. CFU-GM production paralleled
nucleated cell production until day 10, peaked at day 14 (mean 66-fold
increase), and was then maintained until day 21. Cells produced in culture
were predominantly neutrophil precursors and developed normally as assessed
by morphology, immunophenotype, and superoxide generation. This
stroma-free, cytokine-driven culture system can achieve a degree of
amplification, which suggests the feasibility of ex vivo culture of
hematopoietic progenitor cells as an adjunct to hematopoietic stem cell
transplantation.
Volume 80,
Issue 6,
pp. 1405-1412,
09/15/1992
Copyright © 1992 by The American Society of Hematology
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