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Growth-Supporting Activities of Fibronectin on Hematopoietic
Stem/Progenitor Cells In Vitro and In Vivo: Structural Requirement for
Fibronectin Activities of CS1 and Cell-Binding Domains
Takafumi Yokota,
Kenji Oritani,
Hideki Mitsui,
Keisuke Aoyama,
Jun Ishikawa,
Hiroyuki Sugahara,
Itaru Matsumura,
Schickwann Tsai,
Yoshiaki Tomiyama,
Yuzuru Kanakura, and
Yuji Matsuzawa
From the Second Department of Internal Medicine and the Department of
Hematology/Oncology, Osaka University Medical School, Osaka, Japan; and
Fred Hutchinson Cancer Research Center, Seattle, WA.
Fibronectin (FN) is supposed to play important roles in various
aspects of hematopoiesis through binding to very late antigen 4 (VLA4)
and VLA5. However, effects of FN on hematopoietic stem cells are
largely unknown. In an effort to determine if FN had a
growth-supporting activity on hematopoietic stem cells, human CD34+/VLA4bright/VLA5dull
hematopoietic stem cells and a murine stem cell factor (SCF)-dependent multipotent cell line, EML-C1, were treated with or without FN in a
serum and growth-factor-deprived medium, and then subjected to
clonogenic assay in the presence of hematopoietic growth factors. The
pretreatment of the CD34+ cells with FN gave rise to
significantly increased numbers of granulocyte-macrophage
colony-forming units (CFU-GM), erythroid burst colony-forming units,
and mixed erythroid-myeloid colony-forming units. In addition, the
numbers of blast colony-forming units and CFU-GM that developed after
culture of EML-C1 cells with SCF and the combination of SCF and
interleukin-3, respectively, were augmented by the pretreatment with
FN. The augmented colony formation by FN was completely abrogated by
the addition of CS1 fragment, but not of GRGDSP peptide,
suggesting an essential role of FN-VLA4 interaction in the FN effects.
Furthermore, the effects of various FN fragments consisting of
RGDS-containing cell-binding domain (CBD), heparin-binding
domain (HBD), and/or CS1 portion were tested on clonogenic
growth of CD34+ cells. Increased colony formation was
induced by CBD-CS1 and CBD-HBD-CS1 fragments, but not with other
fragments lacking CBD or CS1 domains, suggesting that both CS1 and CBD
of FN were required for the augmentation of clonogenic growth of
hematopoietic stem/progenitor cells in vitro. In addition to the in
vitro effects, the in vivo administration of CBD-CS1 fragment into mice
was found to increase the numbers of hematopoietic progenitor cells in
bone marrow and spleen in a dose-dependent manner. Thus, FN may
function on hematopoietic stem/progenitor cells as a growth-supporting
factor in vitro and in vivo.
Blood, Vol. 91 No. 9 (May 1), 1998:
pp. 3263-3272
© 1998 by The American Society of Hematology.
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