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In Vitro Generation of Megakaryocytes and Platelets from Human Embryonic Stem Cells and Induced Pluripotent Stem Cells

  • Naoya Takayama
  • Koji EtoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 788)

Abstract

Human embryonic stem cells (hESCs) represent a potential source of blood cells for transfusion therapies and a promising tool for studying the ontogeny of hematopoiesis. Moreover, human-induced pluripotent stem cells (hiPSCs), recently established by defined reprogramming factors expressed in somatic cells, represent a further source for the generation of hematopoietic cells. When undifferentiated hESCs or hiPSCs are cultured on either mesenchymal C3H10T1/2 cells or OP-9 stromal cells, they can be differentiated into a hematopoietic niche that concentrates hematopoietic progenitors, which we named “embryonic stem cell-derived sacs” (ES-sacs). We have optimized the in vitro culture condition for obtaining mature megakaryocytes derived from the hematopoietic progenitors within ES-sacs, which are then able to release platelets. These in vitro-generated platelets display integrin activation capability, indicating normal hemostatic function. This novel protocol thus provides a means of generating platelets from hESCs as well as hiPSCs, for the study of normal human thrombopoiesis and also thrombopoiesis in disease conditions using patient-specific hiPSCs.

Key words

Human ESCs Human iPSCs ES-sac Hematopoietic progenitor Megakaryocyte Platelet Transfusion 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  1. 1.Clinical Application Department, Center for iPS Cell Research and Application (CiRA)Kyoto UniversityKyotoJapan
  2. 2.Clinical Application Department, Center for iPS Cell Research and Application (CiRA)Kyoto UniversityKyotoJapan

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