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The Effect of  4 1-Integrin Binding
Sequences of Fibronectin on Growth of Cells From Human Hematopoietic
Progenitors
Karen P. Schofield,
Martin J. Humphries,
Erika de Wynter,
Nydia Testa, and
John T. Gallagher
From the Departments of Medical Oncology and Experimental
Haematology, Paterson Institute for Cancer Research; and the Wellcome
Trust Centre for Cell-Matrix Research, University of Manchester,
Manchester, UK.
Highly regulated interactions between adhesion receptors on
progenitor cells and their extracellular matrix ligands are essential for the control of hematopoiesis in bone marrow stroma. We have examined the relationship between
 4 1-integrin-mediated adhesion and growth
of CD34+ cells by assessing their adhesive and migratory
patterns of proliferation in a mixture of hematopoietic growth factors
in the presence of different recombinant fragments of the
HepII/IIICS region of fibronectin. CD34+ cells
were isolated from cord blood and placed in culture wells containing
serum-free medium and growth factors. Wells were precoated with either
the H120 fragment of fibronectin, which contains three 41-integrin binding sites, or the H0
fragment, which lacks the two highest affinity
41 binding sequences. Proliferation of single cells of CD34+38+DR+
and CD34+38 DR+ phenotypes
occurred in contact with the H120 substrate and was associated with
migration. Larger numbers of cells were used to quantitate
proliferative responses. Cells growing in wells coated with H120 formed
attachments to the base of the wells throughout the culture period.
Higher total cell counts were consistently found in wells coated with
H120 compared with H0 and bovine serum albumin controls.
The difference was first apparent at day 8 of culture and reached a
maximum at days 11 through 13, when expansion with H120 was a mean of
1.8-fold higher than that seen with H0 (P .0001). The
greatest expansion (2.25-fold) with H120 compared with H0 was seen when
the growth factor concentrations were reduced to 1/16 of the standard
levels (P .001). The increase in total cell numbers was not
at the expense of CD34+ cells as numbers of these were
similar in H120 and control cultures. These results provide evidence
for synergy between growth factors and integrins that may be relevant
to understanding hematopoiesis in marrow stroma.
Blood, Vol. 91 No. 9 (May 1), 1998:
pp. 3230-3238
© 1998 by The American Society of Hematology.
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