Mouse embryonic stem cells (mESCs) J1 (strain 129S4/SvJae, RRID:CVCL_6412,) and CJ7 (strain 129S1/SvImJ, RRID:CVCL_C316) were cultured on 0.1% gelatin-coated plates in ESM medium: DMEM supplemented with 15% fetal bovine serum (FBS), 1000 units/mL recombinant leukemia inhibitory factor (LIF), 0.1 mM 2-mercaptoethanol, 2 mM L-glutamine, 0.1 mM MEM non-essential amino acids (NEAA), 1% nucleoside mix (100X stock, Sigma), and 50 U/mL Penicillin/Streptomycin). Primed mouse epiblast stem cells (EpiSCs) were culture on fibronectin-coated plates (10 μg/mL/cm2) in N2B27 medium supplemented with Activin A (20 ng/mL) and Fgf2 (12 ng/mL) (Fidalgo et al., 2016 (link)). Generation of Zfp281KO ESCs (from CJ7 background, clone no. 2.6 (XX), 3.34 (XY) and 7 (XX)) has been previously described (Fidalgo et al., 2011 (link)). All cell lines are from authenticated sources and mycoplasma contamination test was performed routinely.
Free full text: Click here