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. Author manuscript; available in PMC: 2016 Nov 16.
Published in final edited form as: Nat Cell Biol. 2016 May 16;18(6):595–606. doi: 10.1038/ncb3354

Figure 1. Two-step culture of hESCs generates immunophenotypic HSPCs that engraft poorly.

Figure 1

(A) Culture and isolation strategy for differentiating H1 hESCs to HSPCs. (B) Representative FACS plots from 11 experiments staining for CD34, CD90, CD38, CD45 and CD43 on hESC-derived CD34+ cells isolated from 2 week EBs (EB), and after 2 week maturation culture on OP9-M2 (EB-OP9), compared to cells from second trimester foetal liver that were isolated directly (FL) or cultured on OP9-M2 (FL-OP9). (C) Representative FACS plots from 9 experiments staining for CD38, CD34, CD90 and human foetal HSC self-renewal marker GPI-80 on hESC- and FL-derived cells. (D) Human engraftment in NSG mice with hESC-derived and FL-derived CD34+ cells, before and after OP9-M2 co-culture (individual values and mean are shown, n=5 EB, n=4 EB-OP9 and FL, and n=3 FL-OP9 transplanted mice, statistical significance was calculated using the Wilcoxon Rank Sum test, see Supplementary Table 7 for statistics source data). (E) Representative FACS plots of showing human CD45+ fraction in the mouse BM 12 weeks post-transplantation. Multi-lineage engraftment is assessed by CD19 and CD3 (B-and T-lymphoid), and CD66 and CD13 (myeloid) stainings.