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Transendothelial migration of CD34+ and mature hematopoietic cells: an in
vitro study using a human bone marrow endothelial cell line
R Mohle, MA Moore, RL Nachman and S Rafii
Laboratory of Developmental Hematopoiesis, Memorial Sloan-Kettering Cancer
Center, New York, NY 10021, USA.
To study the role of bone marrow endothelial cells (BMEC) in the regulation
of hematopoietic cell trafficking, we have designed an in vitro model of
transendothelial migration of hematopoietic progenitor cells and their
progeny. For these studies, we have taken advantage of a human BMEC-derived
cell line (BMEC-1), which proliferates independent of growth factors, is
contact inhibited, and expresses adhesion molecules similar to BMEC in
vivo. BMEC-1 monolayers were grown to confluency on 3 microns microporous
membrane inserts and placed in 6- well tissue culture plates.
Granulocytecolony stimulating factor (G- CSF)-mobilized peripheral blood
CD34+ cells were added to the BMEC-1 monolayer in the upper chamber of the
6-well plate. After 24 hours of coincubation, the majority of CD34+ cells
remained nonadherent in the upper chamber, while 1.6 +/- 0.3% of the
progenitor cells had transmigrated. Transmigrated CD34 cells expressed a
higher level of CD38 compared with nonmigrating CD34+ cells and may
therefore represent predominantly committed progenitor cells. Accordingly,
the total plating efficiency of the transmigrated CD34+ cells for lineage-
committed progenitors was higher (14.0 +/- 0.1 v 7.8% +/- 1.5%). In
particular, the plating efficiency of transmigrated cells for erythroid
progenitors was 27-fold greater compared with nonmigrating cells (8.0% +/-
0.8% v 0.3% +/- 0.1%) and 5.5-fold compared with unprocessed CD34+ cells
(2.2% +/- 0.4%). While no difference in the expression of the beta
1-integrin very late activation antigen (VLA)-4 and beta 2- integrin
lymphocyte function-associated antigen (LFA)-1 was found, L- selectin
expression on transmigrated CD34+ cells was lost, suggesting that shedding
had occurred during migration. The number of transmigrated cells was
reduced by blocking antibodies to LFA-1, while L-selectin and VLA-4
antibodies had no inhibitory effect. Continuous coculture of the remaining
CD34+ cells in the upper chamber of the transwell inserts resulted in
proliferation and differentiation into myeloid and megakaryocytic cells.
While the majority of cells in the upper chamber comprised proliferating
myeloid precursors such as promyelocytes and myelocytes, only mature
monocytes and granulocytes were detected in the lower chamber. In
conclusion, BMEC-1 cells support transmigration of hematopoietic
progenitors and mature hematopoietic cells. Therefore, this model may be
used to study mechanisms involved in mobilization and homing of CD34+ cells
during peripheral blood progenitor cell transplantation and trafficking of
mature hematopoietic cells.
Volume 89,
Issue 1,
pp. 72-80,
01/01/1997
Copyright © 1997 by The American Society of Hematology
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