T. Budak-alpdogan et al., Morphological and functional characteristics of short-term and long-term bone marrow cultures in chronic myelogenous leukemia, AM J HEMAT, 62(4), 1999, pp. 212-220
Clonogenic capacity of bone marrow progenitors and stromal layers establish
ed from bone marrow of 12 patients with CML and 13 healthy controls were ev
aluated. The initial BFU-E and CFU-GM contents were slightly higher in the
CML patients (p > 0.05) in contrast to CFU-GEMM. CFU-GEMM was lower in the
patients compared to healthy controls (p < 0.001). In long-term cultures, t
he number of non-adherent cell population and total clonogenic progenitor c
ell content decreased gradually in both groups. Weekly evaluation of stroma
l confluency of adherent cells revealed that establishment of adherent stro
mal layer was slower in CML patients than in control samples (p < 0.05). At
the end of fourth week, the number of samples presenting confluency was 41
.7% in the CML group compared with 92.3% in the controls. The initial CD34
positive cell content of the bone marrow samples was similar in both groups
. Although CD34 positive cell number in the adherent stromal layer was well
preserved in the control group at the end of 4 weeks, this figure decrease
d significantly in the CML group. The numbers of total adherent cells as we
ll as the total clonogenic progenitor content of adherent layer were also l
ower in the CML group (3.03% vs 98.2%). When normal CD34+ cells were cultur
ed on IFN-alpha-treated stromal layer followed by the assessment of the lon
g-term culture initiating cells, a reduced capacity to support hemopoietic
growth was observed with IFN-alpha-treated normal stroma. This reduction wa
s even higher when CML stroma was treated with IFN-a followed by the seedin
g of the normal CD34+ cells on this stromal layer (26.9% vs 42.8%). These f
indings show that stromal cells are abnormal in CML patients as well as the
progenitor cells, and IFN-alpha treatment causes further defects of the st
romal cells. (C) 1999 Wiley-Liss, Inc.