Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant

被引:271
作者
Takebe, Takanori [1 ,2 ,3 ]
Zhang, Ran-Ran [1 ]
Koike, Hiroyuki [1 ]
Kimura, Masaki [1 ]
Yoshizawa, Emi [1 ]
Enomura, Masahiro [1 ]
Koike, Naoto [1 ,4 ]
Sekine, Keisuke [1 ]
Taniguchi, Hideki [1 ,2 ]
机构
[1] Yokohama City Univ, Grad Sch Med, Dept Regenerat Med, Yokohama, Kanagawa 232, Japan
[2] Yokohama City Univ, Adv Med Res Ctr, Yokohama, Kanagawa 232, Japan
[3] Japan Sci & Technol Agcy JST, Precursory Res Embryon Sci & Technol, Kawaguchi, Saitama, Japan
[4] Seirei Sakura Citizen Hosp, Dept Surg, Sakura, Japan
关键词
EMBRYONIC STEM-CELLS; DECELLULARIZED MATRIX; ORGANOGENESIS; FIBROBLASTS; MOUSE;
D O I
10.1038/nprot.2014.020
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Generation of functional and vascularized organs from human induced pluripotent stem cells (iPSCs) will facilitate our understanding of human developmental biology and disease modeling, hopefully offering a drug-screening platform and providing novel therapies against end-stage organ failure. Here we describe a protocol for the in vitro generation of a 3D liver bud from human iPSC cultures and the monitoring of further hepatic maturation after transplantation at various ectopic sites. iPSC-derived specified hepatic cells are dissociated and suspended with endothelial cells and mesenchymal stem cells. These mixed cells are then plated onto a presolidified matrix, and they form a 3D spherical tissue mass termed a liver bud (iPSC-LB) in 1-2 d. To facilitate additional maturation, 4-d-old iPSC-LBs are transplanted in the immunodeficient mouse. Live imaging has identified functional blood perfusion into the preformed human vascular networks. Functional analyses show the appearance of multiple hepatic functions in a chronological manner in vivo.
引用
收藏
页码:396 / 409
页数:14
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