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

Frontiers in Cell and Developmental Biology
Francisco Gutierrez-AgüeraPablo Menendez

Abstract

The generation of transplantable hematopoietic stem cells (HSCs) from human pluripotent stem cells (hPSCs) remains challenging. Current differentiation protocols from hPSCs generate mostly hematopoietic progenitors of the primitive HSC-independent program, and it remains unclear what is the best combination of cytokines and hematopoietic growth factors (HGFs) for obtaining functional hematopoietic cells in vitro. Here, we have used the AND1 and H9 hESC lines and the H9:dual-reporter RUNX1C-GFP-SOX17-Cherry to compare the hematopoietic differentiation in vitro based on the treatment of embryoid bodies (EBs) with the ventral mesoderm inducer BMP4 plus HGFs in the absence (protocol 1) or presence (protocol 2) of stage-specific activation of Wnt/β-catenin and inhibition of Activin/Nodal. Despite a slight trend in favor of protocol 1, no statistically significant differences were observed between protocols at any time point analyzed throughout EB development regarding the frequency of hemogenic endothelial (HE) precursors; CD43+ CD45-, CD45+, and CD45 + CD34 + hematopoietic derivatives; or the output of clonogenic progenitors. Similarly, the kinetics of emergence throughout EB development of both SOX17 + HE and RUNX1C + definitive h...Continue Reading

References

Nov 27, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Pablo MenendezMickie Bhatia
Feb 24, 2011·Development·Alexander MedvinskySamir Taoudi
Apr 12, 2012·Molecular Therapy : the Journal of the American Society of Gene Therapy·Pedro J RealPablo Menendez
Sep 5, 2014·Blood·Veronica Ramos-MejíaPablo Menendez
Oct 7, 2015·Methods : a Companion to Methods in Enzymology·Andrea Ditadi, Christopher M Sturgeon
Nov 24, 2016·Nature Reviews. Molecular Cell Biology·Andrea DitadiGordon Keller
Jul 30, 2020·Stem Cells Translational Medicine·Selami DemirciJohn F Tisdale

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