Expandable Megakaryocyte Cell Lines Enable Clinically Applicable Generation of Platelets From Human Induced Pluripotent Stem Cells

Cell Stem Cell. 2014 Apr 3;14(4):535-48. doi: 10.1016/j.stem.2014.01.011. Epub 2014 Feb 13.

Abstract

The donor-dependent supply of platelets is frequently insufficient to meet transfusion needs. To address this issue, we developed a clinically applicable strategy for the derivation of functional platelets from human pluripotent stem cells (PSCs). This approach involves the establishment of stable immortalized megakaryocyte progenitor cell lines (imMKCLs) from PSC-derived hematopoietic progenitors through the overexpression of BMI1 and BCL-XL to respectively suppress senescence and apoptosis and the constrained overexpression of c-MYC to promote proliferation. The resulting imMKCLs can be expanded in culture over extended periods (4-5 months), even after cryopreservation. Halting the overexpression of c-MYC, BMI1, and BCL-XL in growing imMKCLs led to the production of CD42b(+) platelets with functionality comparable to that of native platelets on the basis of a range of assays in vitro and in vivo. The combination of robust expansion capacity and efficient platelet production means that appropriately selected imMKCL clones represent a potentially inexhaustible source of hPSC-derived platelets for clinical application.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Platelets / cytology*
  • Blood Platelets / metabolism
  • Cell Differentiation*
  • Cells, Cultured
  • Disease Models, Animal
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Flow Cytometry
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Induced Pluripotent Stem Cells / metabolism
  • Megakaryocytes / cytology*
  • Megakaryocytes / metabolism
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Polycomb Repressive Complex 1 / metabolism
  • Proto-Oncogene Proteins c-myc / metabolism
  • Thrombocytopenia / metabolism
  • Thrombocytopenia / pathology*
  • bcl-X Protein / metabolism

Substances

  • BCL2L1 protein, human
  • BMI1 protein, human
  • MYC protein, human
  • Proto-Oncogene Proteins c-myc
  • bcl-X Protein
  • Polycomb Repressive Complex 1

Associated data

  • GEO/GSE54168