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Blood, 15 November 2000, Vol. 96, No. 10, pp. 3414-3421
HEMATOPOIESIS
An in vivo competitive repopulation assay for various sources
of human hematopoietic stem cells
Elen S. Rosler,
John E. Brandt,
John Chute, and
Ronald Hoffman
From Hematology-Oncology Section, University of
Illinois College of Medicine, Chicago, IL; Navy-NIDDK Transplantation
and Autoimmunity Branch, Bethesda, MD.
The marrow repopulating potential (MRP) of different sources of
human hematopoietic stem cells (HSCs) was directly compared using an in
vivo assay in which severe combined immunodeficient disease (SCID) mice
were implanted with human fetal bones. HSCs from 2 human lymphocyte
antigen (HLA)-mismatched donors were injected individually or
simultaneously into the fetal bones of a 3rd distinct HLA type and
donor and recipient myeloid and lymphoid cells were identified after 8 to 10 weeks. The study compared the MRP of umbilical cord blood (CB)
and adult bone marrow (ABM) CD34+ cells as well as grafts
of each type expanded ex vivo. Equal numbers of CB and ABM
CD34+ cells injected individually demonstrated similar
abilities to establish multilineage hematopoiesis. However, when CB and
ABM cells were transplanted simultaneously, the engraftment of CB cells
was markedly superior to ABM. CB and ABM CD34+ cells were
expanded ex vivo using either a porcine microvascular endothelial cell
(PMVEC)-based coculture system or a stroma-free expansion system.
Primary CB CD34+ cells or CD34+ cells expanded
in either culture system demonstrated a similar ability to engraft.
However, the MRP of expanded grafts simultaneously injected with
primary CB cells was uniformly inferior to primary CB cells.
CD34+ cell grafts expanded in the stroma-free system,
furthermore, outcompeted CD34+ cells expanded using the
PMVEC coculture system. The triple HLA-mismatched SCID-hu model
represents a novel in vivo stem cell assay system that permits the
direct demonstration of the functional consequences of ex vivo HSC
expansion and ontogeny-related differences in HSCs.
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