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

Published version
Peer-reviewed

Repository DOI


Change log

Authors

Moreau, Thomas 
Evans, Amanda L 
Vasquez, Louella 
Tijssen, Marloes R 
Yan, Ying 

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 × 10(5) 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.

Description

Keywords

Basic Helix-Loop-Helix Transcription Factors, Blood Platelets, Cell Culture Techniques, Cell Differentiation, Cell Proliferation, Cellular Reprogramming, Cryopreservation, GATA1 Transcription Factor, Gene Expression Regulation, Genes, Reporter, Genetic Vectors, Green Fluorescent Proteins, Humans, Lentivirus, Megakaryocytes, Microarray Analysis, Pluripotent Stem Cells, Proto-Oncogene Protein c-fli-1, Proto-Oncogene Proteins, Signal Transduction, T-Cell Acute Lymphocytic Leukemia Protein 1, Transduction, Genetic, Transgenes

Journal Title

Nat Commun

Conference Name

Journal ISSN

2041-1723
2041-1723

Volume Title

7

Publisher

Springer Science and Business Media LLC
Sponsorship
CCF (None)
Medical Research Council (MR/L022982/1)
Medical Research Council (MC_PC_12009)
Medical Research Council (MC_PC_15042)
Wellcome Trust (091310/Z/10/Z)
British Heart Foundation (None)
British Heart Foundation (None)
British Heart Foundation (None)
British Heart Foundation (None)
European Commission (257082)
This work was supported by the Leukemia and Lymphoma Society grant, the UK Medical Research Council (Roger Pedersen), the National Institute for Health Research (NIHR; RP-PG-0310-1002; Willem Ouwehand and Cedric Ghevaert) and a core support grant from the Wellcome Trust and MRC to the Wellcome Trust – Medical Research Council Cambridge Stem Cell Institute. Cedric Ghevaert is supported by the British Heart Foundation (FS/09/039); Marloes Tijssen is supported by the European Hematology Association (Research fellowship) and the British Heart Foundation (PG/13/77/30375). Catherine Hobbs was supported by the National Health Service Blood and Transplant. Matthew Trotter was supported by a Medical Research Council Centre grant (MRC Centre for Stem Cell Biology and Regenerative Medicine); since participation in the work described, Matthew Trotter has become an employee of Celgene Research SLU, part of Celgene Corporation. Nicole Soranzo's research and Sanger Institute affiliates are supported by the Wellcome Trust (WT098051 and WT091310), the EU FP7 (Epigenesys 257082 and Blueprint HEAL TH-F5-2011-282510). The Cambridge Biomedical Centre (BRC) hIPSCs core facility is funded by the NIHR.