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Blood, 15 July 2001, Vol. 98, No. 2, pp. 335-342
HEMATOPOIESIS
Bone morphogenetic protein 4 induces efficient hematopoietic
differentiation of rhesus monkey embryonic stem cells in vitro
Fei Li,
Shijiang Lu,
Loyda Vida,
James A. Thomson, and
George R. Honig
From the Department of Pediatrics, College of Medicine,
University of Illinois at Chicago; and the Wisconsin Regional Primate
Research Center, University of Wisconsin, Madison.
A cell culture system consisting of mouse S17 stromal cells
supplemented with cytokines was developed for hematopoietic
differentiation of rhesus monkey embryonic stem (ES) cells. The
differentiated colonies that formed contained clusters of
hematopoietic-like cells, as well as structures similar in appearance
to embryonic blood islands. When this culture system was supplemented
with bone morphogenetic protein 4 (BMP-4), the numbers of primary
hematopoietic clusters increased by an average of 15 fold. The primary
hematopoietic clusters containing clonogenic precursors (expandable
hematopoietic clusters) increased by 18 fold. Immunofluorescence
analysis showed that a substantial percentage of the
hematopoietic-like cells were CD34+, with morphologic
features of undifferentiated blast cells. Enrichment of the
CD34+ cells was associated with enhanced stromal-dependent,
cytokine-driven formation of cobblestone colonies on secondary plating.
The hematopoietic identity of the precursors was further indicated by
their expression of genes associated with hematopoietic
differentiation, as well as morphologic assessments that showed
erythroid and myeloid lineages among the progeny cells. In addition,
reverse transcriptase-polymerase chain reaction analysis of
BMP-4-treated rhesus monkey ES cells demonstrated an up-regulation of
early-expressed genes responsible for embryonic hematopoiesis and
angiogenesis during the first 7 days of culture. These observations
suggest that embryonic mesoderm regulatory protein may mimic
physiologic signals that are required for the onset of embryonic
hematopoiesis and stem cell formation in rhesus monkey ES cells.
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