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Bone marrow-derived stromal cells prevent apoptotic cell death in B-
lineage acute lymphoblastic leukemia
A Manabe, E Coustan-Smith, FG Behm, SC Raimondi and D Campana
Department of Hematology-Oncology, St Jude Children's Research Hospital,
Memphis, TN 38101.
Establishing requirements for the survival of human immature B cells in
vitro has proved elusive. In this article, we report prolonged survival of
B-lineage leukemic cells on 'feeder layers' of bone marrow (BM)- derived
stromal cells in a serum-free environment. In 15 of 18 cases of B-lineage
acute lymphoblastic leukemia (ALL), there was a greater than 50% decrease
in the number of viable cells after 72 hours of culture in medium alone.
Cell loss was preceded by molecular and cellular changes characteristic of
programmed cell death, or apoptosis, and was not suppressed by adding
interleukin-7 to the tissue culture medium. By contrast, the use of
allogeneic BM stromal cells as feeder layers prevented apoptosis in 10 of
12 cases of ALL, leading to extended survival of the blast cells. This
method was not successful when the allogeneic marrow cells were replaced
with established cell lines. In six of eight cases in which the numbers of
intact CD19+ lymphoblasts were counted by flow cytometry after 7 days of
culture, the proportion of such cells recovered in the presence of BM
stromal cells was 68.8% to 124.7% (median, 95.3%) of that originally
seeded, as opposed to the 0.3% to 15.9% fraction (median, 0.7%) obtained in
the absence of stromal cells. Survival requirements of the B-lineage
lymphoblasts appeared to be heterogeneous, as cells from 3 of the 18 cases
studied showed no signs of apoptosis in serum-free tissue culture medium
that lacked BM stromal cells. The only cells not giving rise to viable
cultures were from two hyperdiploid (greater than 50 chromosomes) cases
with identical karyotypes. The serum-free assay described here can be used
to compare the survival requirements of normal and leukemic B-cell
progenitors as well as to identify the molecules involved in the
interaction between BM stroma and immature B cells.
Volume 79,
Issue 9,
pp. 2370-2377,
05/01/1992
Copyright © 1992 by The American Society of Hematology
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