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R1 mESCs (kindly provided by Dr. Nagy, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada) and mouse STO-SNL2 cells (STO, American Type Culture Collection CRL-2225) were cultivated as previously described48. R1 mESCs were induced to differentiate into embryoid bodies (EBs) by removing the Leukemia Inhibitory Factor from the culture medium (differentiation medium), using the hanging drop method24.
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