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Line VUB02 was derived from IVF embryos at blastocyst stage. Isolated ICMs were transferred to 0.1% gelatin-coated culture dishes containing inactivated mouse embryonic fibroblast (MEF) feeder layers, and cultured further at 37°C in 10% CO2. Karyotype : 46,XY.
VUB02 was derived on inactivated MEFs isolated from 129/Sv mice. The derivation and Culture medium (hES medium): Knockout Dulbecco’s modified Eagle’s medium (KO DMEM) supplemented with fetal bovine serum (FBS, 20%), L-glutamine (2 mM/L), non-essential amino acids (1%), beta-mercaptoethanol (0.1 mmol/l), human recombinant leukaemia inhibitory factor (hrLIF, 103 U/ml) and human recombinant basic fibroblast growth factor (hrbFGF, 4 ng/ml). After a few passages, the culture conditions are changed by using CF1 MEF as feeder cells, omitting hrLIF and replacing FBS with Knockout-Serum Replacement (KO-SR).