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Further phenotypic characterization and isolation of human hematopoietic
progenitor cells using a monoclonal antibody to the c-kit receptor
RA Briddell, VC Broudy, E Bruno, JE Brandt, EF Srour and R Hoffman
Department of Medicine, Indiana University School of Medicine, Indianapolis
46202-5121.
A mouse antihuman monoclonal IgG2a antibody, termed stem cell receptor- 1
(SR-1), specific for a determinant of the c-kit ligand receptor (KR), was
used as an immunologic probe to analyze KR expression by human bone marrow
hematopoietic progenitor cells. Monoclonal antibodies to CD34 and HLA-DR
were used in a multicolor staining protocol in conjunction with SR-1 to
further define the phenotypes of various classes of hematopoietic
progenitor cells. Expression of KR (SR-1+) on hematopoietic progenitor
cells identified subpopulations of cells expressing CD34 (CD34+). While
one-half of the CD34- and HLA-DR- expressing cells (CD34+ HLA-DR+)
expressed the KR (SR-1+), one-third of the CD34+ cells that lacked HLA-DR
expression (CD34+ HLA-DR-) were SR- 1+. The CD34+ HLA-DR+ SR-1+ cell
population contained the vast majority of the more differentiated
progenitor cells, including the colony- forming unit (CFU)
granulocyte-macrophage; burst-forming unit- erythrocyte; CFU-granulocyte,
erythrocyte, macrophage, megakaryocyte; and the CFU-megakaryocyte. The
overall progenitor cell cloning efficiency of this subpopulation was
greater than 31%. By contrast, the CD34+ HLA-DR- SR-1+ cell population
contained fewer of these more differentiated progenitor cells but
exclusively contained the more primitive progenitor cells, the
BFU-megakaryocyte, high proliferative potential-colony-forming cell, and
long-term bone marrow culture- initiating cell. The overall progenitor cell
cloning efficiency of this subpopulation was greater than 7%. Both the
CD34+ HLA-DR- and CD34+ HLA- DR+ cell subpopulations lacking KR expression
contained few assayable hematopoietic progenitor cells. Long-term bone
marrow cultures initiated with CD34+ HLA-DR- SR-1+ but not CD34+ HLA-DR-
SR-1- cells, which were repeatedly supplemented with c-kit ligand (KL) and
interleukin-3, generated assayable progenitor cells of at least 2 lineages
for 10 weeks. These experiments demonstrate the expression of the KR
throughout the hierarchy of human hematopoietic progenitor cell
development. We conclude from our data that the KL and KR play a pivotal
role in cytokine regulation of both the primitive and more differentiated
human hematopoietic progenitor cells.
Volume 79,
Issue 12,
pp. 3159-3167,
06/15/1992
Copyright © 1992 by The American Society of Hematology
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