title: Expandable megakaryocyte cell lines enable clinically applicable generation of platelets from human induced pluripotent stem cells.
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.
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Sou
authors: S
authors: Nakamura
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Naoya
authors: N
authors: Takayama
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Shinji
authors: S
authors: Hirata
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Hideya
authors: H
authors: Seo
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Hiroshi
authors: H
authors: Endo
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Kiyosumi
authors: K
authors: Ochi
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Ken-ichi
authors: K
authors: Fujita
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Tomo
authors: T
authors: Koike
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Ken-ichi
authors: K
authors: Harimoto
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan.
authors: Takeaki
authors: T
authors: Dohda
authors: Department of Reprogramming Science, CiRA, Kyoto University, 606-8507, Japan.
authors: Akira
authors: A
authors: Watanabe
authors: Department of Reprogramming Science, CiRA, Kyoto University, 606-8507, Japan.
authors: Keisuke
authors: K
authors: Okita
authors: Department of Anatomy, Ultrastructural Cell Biology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan.
authors: Nobuyasu
authors: N
authors: Takahashi
authors: Department of Anatomy, Ultrastructural Cell Biology, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan.
authors: Akira
authors: A
authors: Sawaguchi
authors: Department of Reprogramming Science, CiRA, Kyoto University, 606-8507, Japan.
authors: Shinya
authors: S
authors: Yamanaka
authors: Laboratory of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan.
authors: Hiromitsu
authors: H
authors: Nakauchi
authors: Department of Cardiovascular Medicine, The University of Tokyo, Tokyo 113-8655, Japan; Department of Cell and Molecular Medicine, Center for Molecular Medicine, Jichi Medical University, Tochigi 329-0498, Japan.
authors: Satoshi
authors: S
authors: Nishimura
authors: Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, 606-8507, Japan; Laboratory of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, Institute of Medical Science, The University of Tokyo, Tokyo 108-8639, Japan. Electronic address: kojieto@cira.kyoto-u.ac.jp.
authors: Koji
authors: K
authors: Eto
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008533:Megakaryocytes / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D063150:Polycomb Repressive Complex 1 / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism; D013921:Thrombocytopenia / Q000378:metabolism / Q000473:pathology*; D051020:bcl-X Protein / Q000378:metabolism
chemical_list: C494811:BCL2L1 protein, human; C069077:BMI1 protein, human; C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D051020:bcl-X Protein; D063150:Polycomb Repressive Complex 1