autologous blood and marrow transplantation - Blog Science ...

Carlo-Stella et al. 131
DNA fragmentation was detected by terminal deoxynucleotidyl transferase (TdT)
assay. 26
AG957 treatment
AG957 was diluted in dimethylsulfoxide to prepare 1000-fold concentrated
solutions that were used at a final concentration of 0.1% (vol/vol) to obtain the
appropriate concentrations in culture. To evaluate the effect of AG957, CD34 +
cells
(1X 10 5
/mL) were exposed (30 minutes, 37°C, 5% C02) to either control medium
(Iscove's modified Dulbecco's medium [IMDM], 10% FBS) or medium containing
AG957 (1-100 uM). At the end of the incubation period, the cells were washed
three times and cultured to quantitate CFU-Mix, BFU-E, CFU-GM, and LTC-IC.
For each experiment, appropriate controls with vehicle alone (DMSO 1 uL/dish)
were set up.
Statistical analysis
Four plates were scored for each data point per experiment and the results were
expressed as the mean ± 1 standard error of the mean (SEM). Statistical analysis
was performed with the statistical package Statview (Brainpower, Calabasas, CA)
run on a Macintosh 6300 personal computer (Apple, Cupertino, CA). The Student
/ test for paired or unpaired data (two-tail) or the Wilcoxon signed-rank test was
used where appropriate to test the probability of significant differences between
samples. AG957 concentrations resulting in 50% inhibition (ID 50) of colony
formation were calculated for each experiment by extrapolating from a least square
linear regression line relating AG957 concentration to the percentage of colony
growth inhibition.
RESULTS
Effect of AG957 on CML and normal progenitors
As shown in Fig. 1, preincubation of CML CD34 +
cells (n=8) with AG957
(1-100 uM) resulted in a statistically significant (F