Primary AML cells
The study was approved by the local ethics committee (REK Vest) and the Data Inspectorate,
Norway and samples were collected after informed consent in accordance with the
Declaration of Helsinki. Samples were randomly withdrawn from a large group of consecutive
patients with de novo AML and high peripheral blood blast counts [1]. The median age was
64 years (patient data is overviewed in Supplementary table 1 (Table S1)). AML-cells
containing more than 95% tumor cells were prepared using density gradient separation of
peripheral blood samples (Ficoll-Hypaque; NyCoMed, Oslo, Norway; specific density 1.077)
before standardized cryopreservation [2]. The percentage of blasts among leukemic PBMC
exceeded 95% for all patients as judged by light microscopy of May-Grünwald-Giemsa
stained cytospin smears [3-5]. PBMC were cultured in serum free conditions in StemSpan
(Stem Cell Technologies, Vancouver, BC, Canada) at a concentration of 2x106 cells per ml.
1.
2.
3.
4.
5.
6.
Bruserud, O., et al., Flt3-mediated signaling in human acute myelogenous leukemia
(AML) blasts: a functional characterization of Flt3-ligand effects in AML cell
populations with and without genetic Flt3 abnormalities. Haematologica, 2003. 88(4):
p. 416-428.
Bruserud, O., Effect of dipyridamole, theophyllamine and verapamil on spontaneous
in vitro proliferation of myelogenous leukaemia cells. Acta Oncol, 1992. 31(1): p. 538.
Bruserud, O., B.T. Gjertsen, and H.L. von Volkman, In vitro culture of human acute
myelogenous leukemia (AML) cells in serum-free media: studies of native AML blasts
and AML cell lines. J Hematother Stem Cell Res, 2000. 9(6): p. 923-32.
Bruserud, O., et al., Leptin in human acute myelogenous leukemia: studies of in vivo
levels and in vitro effects on native functional leukemia blasts. Haematologica, 2002.
87(6): p. 584-95.
Bruserud, O., et al., Effects of interleukin 10 on blast cells derived from patients with
acute myelogenous leukemia. Leukemia, 1995. 9(11): p. 1910-20.
Mrozek, K., N.A. Heerema, and C.D. Bloomfield, Cytogenetics in acute leukemia.
Blood Rev, 2004. 18(2): p. 115-36.
Table S1. Clinical and biological characteristics of AML samples analysed
Number of patients included
17
Age; median (range)
64 (29-79)
Gender (male/female)
9/8
Predisposition/previous disease
Previous chemotherapy
1
Previous myelodysplasia or chronic myeloproliferative disease
1
AML relapse
2
No predisposition
13
FAB classification
M0/M1
8
M2/M3
1
M4/M5
8
M6/M7
0
Cytogenetic abnormalities
Favourable
1
Normal/intermediate risk
11
High risk
0
Not examined
5
Genetic Flt3 abnormalities
Internal tandem duplication
10/17
Active site mutation
2/17
Wild type
6/17
Genetic Npm1 insertion
5/17
1/15
Genetic CEBP mutations
The cytogenetic abnormalities were classified according to the MRC criteria[6]: (i)
Favourable including inv(16), t(8;21), t(15;17); (ii) high risk including, del(5q), - 7,
inv(3)/t(3;3), - 5, complex; (iii) intermediate, other abnormalities; and (iv) normal karyotype.