Generating Leukemic Npm1c/Flt3-ITD/Cas9 Cells

Npm1c/Flt3-ITD/Cas9 DM cells were generated from lineage-depleted bone marrow cells of primary transgenic mice after leukemia onset (female, 12 weeks old) as described previously^{57 (link),58 (link)}. Cells were maintained in XVIVO-20 medium (Lonza) supplemented with 5% FCS, 1% penicillin-streptomycin-glutamine, mouse SCF 50 ng ml⁻¹ (PeproTech), mouse IL-3 10 ng ml⁻¹ (PeproTech) and mouse IL-6 10 ng ml⁻¹ (R&D Systems), in a 37 °C and 5% CO₂ atmospheric environment. Npm1c/Flt3-ITD/Cas9 DM cells were passaged every 2 days and cultured for a short time (passages 3–5) to maintain the original leukemic properties.

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Lara-Astiaso D., Goñi-Salaverri A., Mendieta-Esteban J., Narayan N., Del Valle C., Gross T., Giotopoulos G., Beinortas T., Navarro-Alonso M., Aguado-Alvaro L.P., Zazpe J., Marchese F., Torrea N., Calvo I.A., Lopez C.K., Alignani D., Lopez A., Saez B., Taylor-King J.P., Prosper F., Fortelny N, & Huntly B.J. (2023). In vivo screening characterizes chromatin factor functions during normal and malignant hematopoiesis. Nature Genetics, 55(9), 1542-1554.

Publication 2023

XVIVO-20 medium penicillin-streptomycin-glutamine mouse IL-3 mouse IL-6

Bone marrow cells Cells Female Flt3 Glutamine Leukemia Mouse Mouse il 6 Penicillin Streptomycin Transgenic mice

Corresponding Organization : University of Salzburg

Other organizations : Universidad de Navarra, Medical Research Council, Wellcome/MRC Cambridge Stem Cell Institute

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Variable analysis

independent variables

Npm1c
Flt3-ITD

dependent variables

Leukemic properties

control variables

Lineage-depleted bone marrow cells of primary transgenic mice after leukemia onset (female, 12 weeks old)
XVIVO-20 medium (Lonza) supplemented with 5% FCS, 1% penicillin-streptomycin-glutamine, mouse SCF 50 ng ml⁻¹ (PeproTech), mouse IL-3 10 ng ml⁻¹ (PeproTech) and mouse IL-6 10 ng ml⁻¹ (R&D Systems)
37 °C and 5% CO₂ atmospheric environment
Short culture time (passages 3–5)

controls

Positive control: Primary transgenic mice after leukemia onset (female, 12 weeks old)
Negative control: Not explicitly mentioned

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