By T. Haferlach, C. Schoch, W. Hiddemann (auth.), Prof. Dr. med. Wolfgang Hiddemann, Prof. Dr. Dr. med. phil. Torsten Haferlach, Dr. med. Michael Unterhalt, Prof. Dr. med. Thomas Büchner, Prof. Dr. med. Jörg Ritter (eds.)
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Extra info for Acute Leukemias IX: Basic Research, Experimental Approaches and Novel Therapies
24 hours later medium was replaced and antibiotics were added (6001200 fig/ml G41S, 150-300 Ilg/ml Zeocin). Dead cells were removed by ficoll gradient 2-6 days later. Individual stably transfected clones were obtained by limiting dilution on 96-well plates. Clones were maintained in 400-600 fig/ml G41S and 100-150 Ilg/ml Zeocin. Analysis of clones 152 clones derived from 12 independent transfections were treated with 1 11M hormone (either Muristerone A or Ponasterone A, Invitrogen, Groningen, NL) and screened for inducible AMLl/ETO protein expression by Western blotting using AMLl- and ETOspecific antibodies (kindly provided by Scott W.
25 Results and Discussion CD34+-progenitor cells were isolated from fresh cord blood at a purity of at least 95%. Virus titer reached up to 1Q6/ml supernatant. The overall transduction efficiency, determined as the ration of colony forming unit (CFU) in the MC assay with or without G418, was 10%. First results indicate that AMLI/ETO transduced cells in liquid culture exhibit a less differentiated morphology with a reduced percentage of granulated cells with prominent vacuolization (Fig. 1, 2). 3).
Yamamoto, C. , Downing, J. , Selsted, M. , and Hiebert, S. W. (1998). The t( 8;21) fusion product, AML-l/ETO, associates with C/EBPa, inhibits C/EBPa-dependent transcription, and blocks granuloctyic differentiation, Mol Cell BioI 18, 322-333. Disturbed Differentiation of AMl1lETO Transduced Human (034+ Progenitors J. BASECKEt, CH. BAUM 2 , O. PFAHLERT l , G. BRITTINGERt, B. WORMANN3, and F. GRIESINGER l Introduction Material and Methods The translocation t( 8;21) (q22;q22), AMLl/ETO, one of the most frequent genetic aberrations in acute myelogenous leukemia is detectable in 40% of AML FAB M2.