西亚试剂：Transient activation of c-MYC expression is critical for ef

Transient activation of c-MYC expression is critical for efficient platelet generation from human induced pluripotent stem cells
Naoya Takayama1, Satoshi Nishimura3,4,5, Sou Nakamura1, Takafumi Shimizu2, Ryoko Ohnishi1, Hiroshi Endo1,2, Tomoyuki Yamaguchi2, Makoto Otsu2, Ken Nishimura4,6, Mahito Nakanishi6, Akira Sawaguchi7, Ryozo Nagai3,5, Kazutoshi Takahashi8, Shinya Yamanaka8, Hiromitsu Nakauchi2, and Koji Eto1

1Stem Cell Bank and 2Division of Stem Cell Therapy, Center for Stem Cell Biology and Regenerative Medicine, the Institute of Medical Science, and 3Department of Cardiovascular Medicine and 5Translational Systems Biology and Medicine Initiative, the University of Tokyo, Tokyo 113-0033, Japan
4PRESTO, Japan Science and Technology Agency, Tokyo 102-8666, Japan
6Gene Function Research Center, National Institute of Advanced Industrial Science and Technology, Ibaraki 305–8562, Japan
7Department of Anatomy, University of Miyazaki Faculty of Medicine, Miyazaki 889-1692, Japan
8Center for iPS Research and Application, Kyoto University, Kyoto 606-8507, Japan

Human (h) induced pluripotent stem cells (iPSCs) are a potentially abundant source of blood cells, but how best to select iPSC clones suitable for this purpose from among the many clones that can be simultaneously established from an identical source is not clear. Using an in vitro culture system yielding a hematopoietic niche that concentrates hematopoietic progenitors, we show that the pattern of c-MYC reactivation after reprogramming influences platelet generation from hiPSCs. During differentiation, reduction of c-MYC expression after initial reactivation of c-MYC expression in selected hiPSC clones was associated with more efficient in vitro generation of CD41a+CD42b+ platelets. This effect was recapitulated in virus integration-free hiPSCs using a doxycycline-controlled c-MYC expression vector. In vivo imaging revealed that these CD42b+ platelets were present in thrombi after laser-induced vessel wall injury. In contrast, sustained and excessive c-MYC expression in megakaryocytes was accompanied by increased p14 (ARF) and p16 (INK4A) expression, decreased GATA1 expression, and impaired production of functional platelets. These findings suggest that the pattern of c-MYC expression, particularly its later decline, is key to producing functional platelets from selected iPSC clones.

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