Mouse MLL-AF9 leukemic cells with Dot1lfl/fl or wild-type genotypes were generated by transformation of mouse bone marrow Lin−Sca1+cKit+ (LSK) cells with retrovirus expressing MLL-AF9 fusion protein and transplanted into sublethally irradiated recipient mice as described previously8 (link). The leukemic blasts harvested from the diseased mice were cultured in vitro in IMDM plus 15% FBS supplemented with 20 ng/ml murine SCF (PeproTech), 10 ng/ml murine IL-3 (PeproTech) and 10 ng/ml murine IL-6 (PeproTech). Human leukemic cell lines Molm-13, MV4-11 and HL-60 were maintained in RPMI plus 10% FBS. All cell culture medium contained L-Glutamine (2mM; Gibco), penicillin (100 units/ml; Gibco), streptomycin (100 ug/ml; Gibco) and plasmocin (5 ug/ml; InvivoGen). Human cell lines including HL-60, MV4-11 and Molm-13 were tested in March - May, 2013 for authentication by short tandem repeat (STR) profiling performed by ATCC. Live cell counts were obtained by high-throughput flow cytometry with SYTOX-blue cell death dye exclusion (Life Technologies) using an LSRFortessa-HTS Analyzer (BD biosciences).
Sytox blue cell death dye exclusion
Manufactured by Thermo Fisher Scientific
SYTOX-blue cell death dye exclusion is a laboratory reagent used to detect cell viability. It functions by selectively staining dead or dying cells, allowing for the differentiation between viable and non-viable cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Murine scf, by Thermo Fisher Scientific (2 mentions)
Murine il 3, by Thermo Fisher Scientific (2 mentions)
Murine il 6, by Thermo Fisher Scientific (2 mentions)
Plasmocin, by InvivoGen (2 mentions)
Lsrfortessa hts analyzer, by BD (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
2 protocols using sytox blue cell death dye exclusion
1
Murine MLL-AF9 Leukemia Model
2
Murine MLL-AF9 Leukemia Model
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Mouse MLL-AF9 leukemic cells with Dot1lfl/fl or wild-type genotypes were generated by transformation of mouse bone marrow Lin−Sca1+cKit+ (LSK) cells with retrovirus expressing MLL-AF9 fusion protein and transplanted into sublethally irradiated recipient mice as described previously8 (link). The leukemic blasts harvested from the diseased mice were cultured in vitro in IMDM plus 15% FBS supplemented with 20 ng/ml murine SCF (PeproTech), 10 ng/ml murine IL-3 (PeproTech) and 10 ng/ml murine IL-6 (PeproTech). Human leukemic cell lines Molm-13, MV4-11 and HL-60 were maintained in RPMI plus 10% FBS. All cell culture medium contained L-Glutamine (2mM; Gibco), penicillin (100 units/ml; Gibco), streptomycin (100 ug/ml; Gibco) and plasmocin (5 ug/ml; InvivoGen). Human cell lines including HL-60, MV4-11 and Molm-13 were tested in March - May, 2013 for authentication by short tandem repeat (STR) profiling performed by ATCC. Live cell counts were obtained by high-throughput flow cytometry with SYTOX-blue cell death dye exclusion (Life Technologies) using an LSRFortessa-HTS Analyzer (BD biosciences).
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