Robust generation of erythroid and multilineage hematopoietic progenitors from human iPSCs using a scalable monolayer culture system

作者：Juan Pablo Ruíz, Guibin Chen, Juan J. Haro‐Mora, Keyvan Keyvanfar, Chengyu Liu, Jizhong Zou, Jeanette Beers, Hanan Bloomer, Husam Qanash, Naoya Uchida, John F. Tisdale, Manfred Boehm, André Larochelle · 发表于：Stem Cell Research · 年份：2019 · DOI：10.1016/j.scr.2019.101600 · 被引用次数：48 · 研究领域：Zebrafish Biomedical Research Applications、Pluripotent Stem Cells Research、Epigenetics and DNA Methylation

One of the most promising objectives of clinical hematology is to derive engraftable autologous hematopoietic stem cells (HSCs) from human induced pluripotent stem cells (iPSCs). Progress in translating iPSC technologies to the clinic relies on the availability of scalable differentiation methodologies. In this study, human iPSCs were differentiated for 21 days using STEMdiff™, a monolayer-based approach for hematopoietic differentiation of human iPSCs that requires no replating, co-culture or embryoid body formation. Both hematopoietic and non-hematopoietic cells were functionally characterized throughout differentiation. In the hematopoietic fraction, an early transient population of primitive CD235a+ erythroid progenitor cells first emerged, followed by hematopoietic progenitors with multilineage differentiation activity in vitro but no long-term engraftment potential in vivo. In later stages of differentiation, a nearly exclusive production of definitive erythroid progenitors was observed. In the non-hematopoietic fraction, we identified a prevalent population of mesenchymal stromal cells and limited arterial vascular endothelium (VE), suggesting that the cellular constitution of the monolayer may be inadequate to support the generation of HSCs with durable repopulating potential. Quantitative modulation of WNT/β-catenin and activin/nodal/TGFβ signaling pathways with CHIR/SB molecules during differentiation enhanced formation of arterial VE, definitive multilineage and er...