A Benchmark Side-by-Side Comparison of Two Well-Established Protocols for in vitro Hematopoietic Differentiation From Human Pluripotent Stem Cells

The generation of transplantable hematopoietic stem cells (HSCs) from human pluripotent (hPSCs) remains challenging. Current differentiation protocols hPSCs generate mostly progenitors the primitive HSC-independent program, and it unclear what is best combination cytokines growth factors (HGFs) for obtaining functional in vitro . Here, we have used AND1 H9 hESC lines H9:dual-reporter RUNX1C -GFP- SOX17 -Cherry to compare based on treatment embryoid bodies (EBs) with ventral mesoderm inducer BMP4 plus HGFs absence (protocol 1) or presence 2) stage-specific activation Wnt/β-catenin inhibition Activin/Nodal. Despite a slight trend favor protocol 1, no statistically significant differences were observed between at any time point analyzed throughout EB development regarding frequency hemogenic endothelial (HE) precursors; CD43+ CD45−, CD45+, CD45 + CD34 derivatives; output clonogenic progenitors. Similarly, kinetics emergence both HE definitive hematopoiesis was very similar protocols. expression early master mesendodermal transcription Brachyury, MIXL1, KDR revealed gene prior Collectively, simpler 1 is, least, as efficient 2, suggesting that supplementation additional morphogens/HGFs modulation Activin/Nodal pathways seem dispensable hPSCs.