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

T Takebe, RR Zhang, H Koike, M Kimura… - Nature protocols, 2014 - nature.com
T Takebe, RR Zhang, H Koike, M Kimura, E Yoshizawa, M Enomura, N Koike, K Sekine
Nature protocols, 2014nature.com
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 …
Abstract
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.
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