(A) Strategy for comparing the expansion of hESC- and FL-HSPCs. (B) FACS staining for HSPC surface markers CD34+, CD38−/lo, CD45+ and CD90+ at various time points in OP9-M2 co-culture. (C) Expansion of FL and hESC-derived haematopoietic cells sorted for HSPC phenotype after two-step differentiation, and cultured for additional weeks on OP9-M2 (mean +/- SEM – upward bars – from n=4 experiments, statistical significance was assessed using the Wilcoxon Rank Sum test. (D) CFU-C expansions from 10,000 hESC-derived or FL-derived immunophenotypic HSPCs in methylcellulose following 0, 1 and 3 additional weeks on OP9-M2 co-culture (mean +/- SEM – upward bars –from n=4 experiments, statistical significance was assessed using the Wilcoxon Rank Sum test. (E) The morphology of myelo-erythroid colonies generated from hESC- or FL-HSPCs on methylcellulose as assessed by light microscopy (green scale bar = 1mm) and May-Grünwald-Giemsa (MGG) staining (black scale bar = 100 μm). Statistics source data used to generate graphs in C and D can be found in Supplementary Table 7.