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. 2014 May 16;166(3):435–448. doi: 10.1111/bjh.12910

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© 2014 The Authors. British Journal of Haematology published by John Wiley & Sons Ltd.

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Multi-lineage haemopoietic differentiation of hiPSC lines upon OP9 co-culture. (A) Human induced pluripotent stem cells (hiPSC) lines from a variety of sources were assessed for their haemopoietic potential. hiPSC colonies were harvested and co-cultured on OP9 stroma for 10 d. Haemopoietic progeny arising after 10 d co-culture with OP9 were harvested by collagenase/trypsin treatment and analysed for the appearance of markers typical of haemogenic endothelium committed to the haemopoietic lineage that includes staining for CD34⁺ with CD43⁺, and CD43⁺ with CD144⁺ and CD31⁺. CD34⁺ cells were isolated by magnetic-activated cell separation, and seeded onto MS5 feeders and exposed to either (B) CD146⁺ networks, (C) lymphoid (SCF, Flt3L, IL7 and IL3) or (D) erythroid (SCF, EPO, IGF, IL3 and dexamethasone) conditions for a further 21 d. Here we see the appearance of either CD19⁺CD10⁺ B lymphocytes or CD71⁺ CD235a⁺ erythroblasts, as well as CD146⁺ endothelial networks, indicating that all hiPSC lines differentiate into a multi-lineage progenitor that is capable of haemato-endothelial differentiation. Mean data are provided in the far right panels where n = 2–3, and standard deviation is shown.