Illumina HiSeq 4000 paired end sequencing; Primordial germ cell like cell competent populations
Primordial germ cell like cell competent populations
ME_p
hESCs, PreME and ME cells were dissociated using TrypLE, neutralised using MEF media and sorted to remove doublets and debries using a SONY SH800 sorter hESCs were cultured using E8 media on vitronectin coated plates. PreME and ME cells were grown according to Kobayashi et al 2017. PreME/ME protocols were optimised based on previous conditions. For the basal medium, we used aRB27 medium which is composed of Advanced RPMI 1640 Medium (Thermo Fisher Scientific) supplemented with 1% B27 supplement (Thermo Fisher Scientific), 0.1 mM NEAA, 100 U/ml Penicillin- 0.1 mg/ml Streptomycin, 2 mM L-Glutamine. For PreME/ME induction, trypsinized Conv-hPSC were seeded on vitronectin-coated dish at 200,000 cells/well in 12 well plate and cultured in PreME/ME induction medium for 6-36 h as indicated in the figures.  PreME/ME induction medium was composed of aRB27 medium supplemental with 100 ng/ml Activin-A (SCI), 3 M GSK3i and 10 M of ROCKi.   Single Cell 3' 10X V2  Single Cell 3' 10X V2