organism: Mus musculus
attributes.tag: isolate
attributes.value: DIV5_KO90_KO_2
attributes.tag: breed
attributes.value: 129/Ola
attributes.tag: dev_stage
attributes.value: 5 DIV
attributes.tag: collection_date
attributes.value: not applicable
attributes.tag: geo_loc_name
attributes.value: not applicable
attributes.tag: sex
attributes.value: male
attributes.tag: tissue
attributes.value: blastocyst
attributes.tag: cell_line
attributes.value: E14Tg2a
attributes.tag: cell_type
attributes.value: Mettl9 KO
attributes.tag: growth_protocol
attributes.value: mESCs were cultured in Serum/LIF (Dulbecco′s Modified Eagle′s Medium - high glucose), 15% Fetal bovine serum, 2 mM L-Glutamine, 1X MEM Non-Essential Amino Acids Solution, 1 mM Sodium Pyruvate, 50 μM 2-Mercaptoethanol, 5 ng/ml (1000U/ml) LIF. mESCs were detached with Trypsin-EDTA 1X or StemPro™ Accutase™ Cell Dissociation Reagent and seeded on Gelatin-coated plates (EmbryoMax® 0.1% Gelatin Solution). For neural differentiation, mESC were cultured in 2i/LIF (DMEM/F-12, no glutamine, 1X B-27™ Supplement, minus vitamin A, 1X N-2 Supplement, 0.1X MEM Non-Essential Amino Acids Solution, 2 mM L-Glutamine, 1 mM Sodium Pyruvate, 50 μM 2-Mercaptoethanol, 1 μM PD0325901- MEK inhibitor , 3 μM CHIR99021 - GSK3 inhibitor, for 1 day (DIV-1 > DIV0). After 24 hrs (DIV0), 2i/LIF media was replaced by WiBi media (DMEM/F-12, no glutamine, 1X B-27™ Supplement (50X), minus vitamin A #12587010, 1X N-2 Supplement (100X) # 17502048, 0.1X MEM Non-Essential Amino Acids Solution, 2 mM L-Glutamine, 1 mM Sodium Pyruvate, 50 μM 2-Mercaptoethanol, 2.5 μM 54AH-Wnt pathway inhibitor, 0.2 μM LDN193189 hydrochloride, which was provided fresh daily until DIV10 (endpoint). From DIV5 until DIV10, WiBi media was supplemented with 0.1 μM SAG dihydrochloride to induce a basal telencephalic fate. During neural differentiation (from DIV3 onwards), cells were cultured on Poly-L-ornithine and Laminin coated plates, with 16 μg/ml Poly-L-ornithine hydrobromide diluted in H20 and 3.2 μg/ml Laminin Mouse Protein diluted in PBS. All cells were cultured at 37 °C with 5% CO2.
attributes.tag: BioSampleModel
attributes.value: Model organism or animal