12th Congress of the European Hematology ... - Haematologica

12 th Congress of the European Hematology Association
with de novo MDS, who received an allograft using fludarabine/busulfex
or fludarabine/busulfex/antithymocyte globulin (ATG). Results. Nineteen
patients (86.4%) achieved engraftment. At a median follow-up of 18.9
months (range, 13.1-24.8 months), the estimated 2-year overall survival
(OS), event-free survival (EFS), transplantation-related mortality, and
relapse incidence were 78.7, 67.7, 12.6, and 22.5%, respectively. Acute
graft-versus-host disease (GVHD) greater than grade II developed in three
patients (15.8%). Chronic GVHD developed in 10 patients (55.6%) and
none of these patients received ATG as a conditioning regimen. Variables
influencing EFS were chronic GVHD, marrow blasts before transplantation,
and the WHO criteria. Conclusions. The present study clarifies
the benefits of the fludarabine/busulfex-based conditioning regimen for
de novo MDS diagnosed according to the WHO criteria, and shows that
chronic GVHD appears to have a beneficial effect on survival rates, which
are strongly associated with graft-versus-tumor effects.
358 | haematologica/the hematology journal | 2007; 92(s1)
0962
STUDY OF APOPTOSIS AND EXPRESSION PATTERN OF TNF RECEPTOR FAMILY MEMBERS
IN HUMAN BONE MARROW MESECHYMAL STEM CELLS
H. Papadaki, V.M. Vlahava, M.C. Kastrinaki, A. Damianaki,
C. Gemetzi, G.D. Eliopoulos
University of Crete School of Medicine, VOUTES, HERAKLION, Greece
Background. There is currently great interest in exploring the use of
bone marrow (BM) derived mesenchymal stem cells (MSCs) for the repair
of tissue damage. The expression of tumor necrosis factor receptor
(TNFR) family members and survival characteristics of MSCs in inflammatory
microenvironment is entirely unknown Aims. The aim of the
study was to investigate (a) the expression of the apoptosis-related TNFR
family members in human BM MSCs and (b) the apoptotic characteristics
of BM MSCs in standard culture conditions and following incubation
with TNF family members. Methods. Normal human BM cells were isolated
from posterior iliac crest aspirates and MSCs were expanded according
to a standard protocol. MSC identification was based on immunophenotypic
characteristics (CD45- , CD14 – , CD34 – , CD90 + , CD73 + , CD44 + ,
CD29 + , CD105 + , CD146 + ) and the potential of cells to differentiate
towards the adipogenic (Oil red O stain and aP2 and PPAR-Á expression
by RT-PCR), osteogenic (ALP/Von Kossa stain and ALP and CBFA1
expression by RT-PCR) and chondrogenic (Masson and Alcian blue stain
and Collagen II and Aggrecan expression by RT-PCR) lineages. Flowcytometry
and RT-PCR was used for evaluation of the expression of the
apoptosis-related TNFR family members namely Fas, TNFRI, TNFRII,
TNF related apoptosis-inducing ligand (TRAIL) R1, TRAILR2, TRAILR3,
TRAILR4, time-course from Passage (P) 1-6 The survival characteristics
of MSCs under different culture conditions were evaluated using flowcytometry
and 7-aminoactinomycin D (7AAD) stain. Results. Spontaneous
apoptosis of MSCs at P2 was elevated to 10.02%±6.48% and this
percentage remained stable time-course. As expected, the presence of
10% fetal calf serum (FCS) in the culture medium protected MSCs from
spontaneous apoptosis compared to FCS deprived cultures (p