Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant.
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.
1] Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan. [2] Advanced Medical Research Center, Yokohama City University, Yokohama, Japan. [3] Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Kawaguchi, Japan.
Takanori
T
Takebe
1] Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan. [2].
Ran-Ran
RR
Zhang
1] Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan. [2].
Hiroyuki
H
Koike
1] Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan. [2].
Masaki
M
Kimura
1] Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan. [2].
Emi
E
Yoshizawa
Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
Masahiro
M
Enomura
1] Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan. [2] Department of Surgery, Seirei Sakura Citizen Hospital, Ebaradai, Sakura, Japan.
Naoto
N
Koike
Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
Keisuke
K
Sekine
1] Department of Regenerative Medicine, Yokohama City University Graduate School of Medicine, Yokohama, Japan. [2] Advanced Medical Research Center, Yokohama City University, Yokohama, Japan.
Hideki
H
Taniguchi
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008099:Liver / Q000254:growth & development*; D016031:Liver Transplantation / Q000379:methods; D051379:Mice; D038081:Organogenesis / Q000502:physiology*; D044968:Regenerative Medicine / Q000379:methods*; D033581:Stem Cell Transplantation / Q000379:methods*; D046509:Tissue Culture Techniques