Efficient production of erythroid, megakaryoid and myeloid cells, using single cell-derived iPSC colony differentiation

Abstract

Hematopoietic differentiation of human induced pluripotent stem cells (iPSCs) provide opportunities not only for fundamental research and disease modelling/drug testing but also for large-scale production of blood effector cells for future clinical application. Although there are multiple ways to differentiate human iPSCs towards hematopoietic lineages, there is a need to develop reproducible and robust protocols. Here we introduce an efficient way to produce three major blood cell types using a standardized differentiation protocol that starts with a single hematopoietic initiation step. This system is feeder-free, avoids EB-formation, starts with a hematopoietic initiation step based on a novel single cell-derived iPSC colony differentiation and produces multi-potential progenitors within 8-10 days. Followed by lineage-specific growth factor supplementation these cells can be matured into well characterized erythroid, megakaryoid and myeloid cells with high-purity, without transcription factor overexpression or any kind of pre-purification step. This standardized differentiation system provides a simple platform to produce specific blood cells in a reproducible manner for hematopoietic development studies, disease modelling, drug testing and the potential for future therapeutic applications.\n\nHighlightsO_LIEfficient hematopoietic differentiation from single cell-derived iPSC colonies\nC_LIO_LIReproducible feeder-free, monolayer differentiation system independent of iPSC line\nC_LIO_LIProduction of erythroid, megakaryoid and myeloid cells with high-purity\nC_LIO_LIPlatform for hematopoietic developmental research and future clinical application\nC_LI