Feeder-free Human ESC Maintenance and Differentiation

Human ESC lines were maintained on human foreskin fibroblast feeders or in feeder-free culture conditions on Matrigel (BD Biosciences) in mTeSR1 medium (STEMCELL Technologies) as previously described (Narva et al., 2012 (link), Konki et al., 2016 (link)). In feeder-free culture conditions the cells were maintained on Matrigel (BD Biosciences) in mTeSR1 medium (STEMCELL). Differentiation of hESCs was performed as described by Narva et al. (2012) (link). In brief, for spontaneous embryonic body differentiation the cells were plated without feeders and were grown in suspension in standard hESC medium without fibroblast growth factor 2. For retinoic acid-induced differentiation, cells were plated in feeder-free conditions and medium was supplemented with 13.7 μM retinoic acid (Sigma). The karyotypes of the lines were routinely monitored with G-banding and/or KaryoLite BoBs assay (Lund et al., 2012 (link)).