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Lymphoid and myeloid differentiation of single human CD34+, HLA-DR+, CD38-
hematopoietic stem cells
S Huang and LW Terstappen
Becton Dickinson Immunocytometry Systems, San Jose, CA 95131.
Multilineage differentiation of human fetal bone marrow CD34+ cell subsets
was examined using a single-cell liquid culture assay. Four CD34+ cell
populations, ie, (1) CD38-, HLA-DR+, (2) CD38-, HLA-DR-, (3) CD38+,
HLA-DR-, and (4) CD38+, HLA-DR+ cells, were sorted as single cells into
96-well flat-bottom culture plates containing long-term culture medium
supplemented with interleukin-3, interleukin-6, stem cell factor (SCF),
granulocyte-macrophage colony-stimulating factor, erythropoietin, basic
fibroblast growth factor (bFGF), and insulin-like growth factor-1 (IGF-1).
Single CD34+, CD38-, HLA-DR+ cells had the highest replating efficiency as
well as the highest replating efficiency. The cellular composition of the
single-cell progeny was studied by morphologic and/or flow cytometric
examination. Only the progeny of single CD34+ cells that lacked CD38 could
give rise to each of the hematopoietic cell lineages. The expansion of the
progeny of single CD34+, CD38-, HLA-DR+ cells was examined in more detail
and showed three clearly distinguishable growth patterns: 28% (SD, +/- 10%;
n = 14) of the single cells formed cell clusters/colonies; 9% (SD, +/- 4%;
n = 14) formed dispersed cells; and 11% (SD, +/- 6%; n = 14) gave rise to a
mixture of cell clusters and dispersed cells. The dispersed cell growth
pattern was reduced when SCF or bFGF and IGF-1 was absent in the growth
factor cocktail. The replating ability of the dispersed cells was
considerably larger than that of cells with other growth patterns, in that
76% of the cells that gave rise to dispersed cells and 54% of the cells
that gave rise to dispersed cells as well as cell clusters gave rise to a
second generation, but only 7% of the cells that gave rise to cell clusters
gave rise to a second generation. The second generation of cells continued
to produce third and fourth generations after repetitive replating, except
for the replated cells from cell clusters. In contrast with the
first-generation progeny, SCF did not have an influence on the replating
ability of the cells. Only in the progeny of single CD34+, CD38-, HLA-DR+
cells that gave rise to dispersed cells was each of the hematopoietic cell
lineages found, ie, B lymphocytes, neutrophils, monocytes, macrophages,
osteoclasts, basophils/mast cells, eosinophils, erythrocytes,
megakaryocytes, and platelets.(ABSTRACT TRUNCATED AT 400 WORDS)
Volume 83,
Issue 6,
pp. 1515-1526,
03/15/1994
Copyright © 1994 by The American Society of Hematology
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