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

Nat Commun. 2016 Apr 7:7:11208. doi: 10.1038/ncomms11208.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • FLI1 protein, human
  • GATA1 Transcription Factor
  • GATA1 protein, human
  • Proto-Oncogene Protein c-fli-1
  • Proto-Oncogene Proteins
  • T-Cell Acute Lymphocytic Leukemia Protein 1
  • TAL1 protein, human
  • Green Fluorescent Proteins