Large-scale production of megakaryocytes from human pluripotent stem cells by chemically defined forward programming

Thomas Moreau, Amanda L. Evans, Louella Vasquez, Marloes R. Tijssen, Ying Yan, Matthew W. Trotter, Daniel Howard, Maria Colzani, Meera Arumugam, Wing Han Wu, Amanda Dalby, Riina Lampela, Guenaelle Bouet, Catherine M. Hobbs, Dean C. Pask, Holly Payne, Tatyana Ponomaryov, Alexander Brill, Nicole Soranzo, Willem H. Ouwehand, Roger A. Pedersen () and Cedric Ghevaert ()
Additional contact information
Thomas Moreau: University of Cambridge and NHS Blood and Transplant
Amanda L. Evans: University of Cambridge and NHS Blood and Transplant
Louella Vasquez: Human Genetics, Wellcome Trust Sanger Institute, Genome Campus
Marloes R. Tijssen: University of Cambridge and NHS Blood and Transplant
Ying Yan: Human Genetics, Wellcome Trust Sanger Institute, Genome Campus
Matthew W. Trotter: The Anne McLaren Laboratory, University of Cambridge
Daniel Howard: University of Cambridge and NHS Blood and Transplant
Maria Colzani: University of Cambridge and NHS Blood and Transplant
Meera Arumugam: University of Cambridge and NHS Blood and Transplant
Wing Han Wu: University of Cambridge and NHS Blood and Transplant
Amanda Dalby: University of Cambridge and NHS Blood and Transplant
Riina Lampela: Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute
Guenaelle Bouet: University of Cambridge and NHS Blood and Transplant
Catherine M. Hobbs: University of Cambridge and NHS Blood and Transplant
Dean C. Pask: University of Cambridge and NHS Blood and Transplant
Holly Payne: Institute of Cardiovascular Sciences, University of Birmingham
Tatyana Ponomaryov: Institute of Cardiovascular Sciences, University of Birmingham
Alexander Brill: Institute of Cardiovascular Sciences, University of Birmingham
Nicole Soranzo: Human Genetics, Wellcome Trust Sanger Institute, Genome Campus
Willem H. Ouwehand: University of Cambridge and NHS Blood and Transplant
Roger A. Pedersen: The Anne McLaren Laboratory, University of Cambridge
Cedric Ghevaert: University of Cambridge and NHS Blood and Transplant

Nature Communications, 2016, vol. 7, issue 1, 1-16

Abstract: Abstract The production of megakaryocytes (MKs)—the precursors of blood platelets—from human pluripotent stem cells (hPSCs) offers exciting clinical opportunities for transfusion medicine. Here we describe an original approach for the large-scale generation of MKs in chemically defined conditions using a forward programming strategy relying on the concurrent exogenous expression of three transcription factors: GATA1, FLI1 and TAL1. The forward programmed MKs proliferate and differentiate in culture for several months with MK purity over 90% reaching up to 2 × 105 mature MKs per input hPSC. Functional platelets are generated throughout the culture allowing the prospective collection of several transfusion units from as few as 1 million starting hPSCs. The high cell purity and yield achieved by MK forward programming, combined with efficient cryopreservation and good manufacturing practice (GMP)-compatible culture, make this approach eminently suitable to both in vitro production of platelets for transfusion and basic research in MK and platelet biology.

Date: 2016
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DOI: 10.1038/ncomms11208

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