TY - JOUR
T1 - Assembly and Function of a Bioengineered Human Liver for Transplantation Generated Solely from Induced Pluripotent Stem Cells
AU - Takeishi, Kazuki
AU - Collin de l'Hortet, Alexandra
AU - Wang, Yang
AU - Handa, Kan
AU - Guzman-Lepe, Jorge
AU - Matsubara, Kentaro
AU - Morita, Kazutoyo
AU - Jang, Sae
AU - Haep, Nils
AU - Florentino, Rodrigo M.
AU - Yuan, Fangchao
AU - Fukumitsu, Ken
AU - Tobita, Kimimasa
AU - Sun, Wendell
AU - Franks, Jonathan
AU - Delgado, Evan R.
AU - Shapiro, Erik M.
AU - Fraunhoffer, Nicolas A.
AU - Duncan, Andrew W.
AU - Yagi, Hiroshi
AU - Mashimo, Tomoji
AU - Fox, Ira J.
AU - Soto-Gutierrez, Alejandro
N1 - Funding Information:
De-identified tissues were obtained from Magee Women’s Hospital (Pittsburgh, PA) and the University of Washington Department of Pediatrics, Division of Genetic Medicine, Laboratory of Developmental Biology (Seattle, WA) after obtaining a written informed consent by a protocol approved by the Human Research Review Committee of the University of Pittsburgh (Honest broker approval number HB015 and HB000836). Human fetal hepatocytes were isolated and culture from fetal livers as previously described ( Tobita et al., 2016 ). The de-identified normal human liver cells were obtained through the Liver Tissue Cell Distribution System (Pittsburgh, PA) after obtaining a written informed consent by a protocol approved by the Human Research Review Committee of the University of Pittsburgh, which was funded by NIH Contract # HSN276201200017C. Adult human hepatocytes were also obtained from Ira J Fox Laboratory at Children’s Hospital of UPMC, after obtaining a written informed consent by a protocol approved by the Human Research Review Committee and the Institutional Review Board (IRB#: PRO12090466) of the University of Pittsburgh. For human fetal or adult liver tissue and hepatocytes used in this study both genders were used (XX and XY) and the ages for fetal livers ranged from 16 to 24 weeks of gestational age and for adult livers ages ranged from 32yo to 52yo. Specific information on age, gender and cell viability of human liver tissue and hepatocytes used in this study is described in Table S2 .
Funding Information:
We thank Stephen C. Strom for sharing human iPSCs that have been used in part of this report. We also thank the Center for Biologic Imaging at the University of Pittsburgh for technical support on the transmission electro microscopy study. This project used the UPMC Hillman Cancer Center and Tissue and Research Pathology/Pitt Biospecimen Core shared resource, which is supported in part by award P30CA047904. This project was also partially supported by the Pilot and Feasibility program under the NIH/NIDDK Digestive Disease Research Core Center grant P30DK120531. This work was supported by NIH grants DK099257, DK117881, DK119973, and TR002383 to A.S.-G. P01DK096990 to I.J.F. and A.S.G. DK107697 to E.M.S. and A.S.G. and DK103645 to A.W.D. and by the American Liver Foundation to K.T. K.T. and A.S.-G. conceived and designed the study; K.T. A.C.H. K.H. Y.W. J.G.-L. N.A.F. K.M. S.J. J.F. E.R.D. A.W.D. H.Y. and A.S.-G. performed data acquisition; K.T. A.C.H. K.H. Y.W. J.G.-L. S.J. N.H. R.M.F. A.W.D. H.Y. I.J.F. and A.S.-G. analyzed and interpreted data; K.F. K.H. J.G.-L. and W.S. performed rat liver decellularization and characterization; K.F. K.T. and E.M.S. developed and performed iron-micro-particles for imaging studies to evaluate organ assembly; K.T. A.C.H. K.H. J.G.-L. and A.S.-G. established and performed hepatic differentiation of human iPSCs; Y.W. K.T. F.Y. J.G.-L. and A.S.-G. established and performed cholangiocyte differentiation of human iPSCs; K.T. A.C.H. Y.W. J.G.-L. and S.J. performed rat liver assembly and characterization; Y.W. and K.M. established an auxiliary liver transplantation model and performed rat liver procurement and transplantation; T.M. established the IL2rg?/? rat; K.T. I.J.F. and A.S.-G. wrote the manuscript; K.T. A.C.H. H.Y. I.J.F. and A.S.-G. participated in critical revision of the manuscript for intellectual content; and A.S.-G. obtained funding. All authors contributed to the preparation of the manuscript. H.Y. and A.S.-Gare inventors on a patent application that involves some of the perfusion technology used in this work (WO/2011/002926); K.H. K.M. J.G.-L. H.Y. and A.S.-G. have an international patent related to this work that describes methods of preparing artificial organs and related compositions for transplantation and regeneration (WO/2015/168254). K.T. A.C.H. J.G.-L. Y.W. T.M. and A.S.-G. have a provisional international patent application that describes hepatic differentiation of human pluripotent stem cells and liver repopulation (PCT/US2018/018032). A.S.-G. J.G.-L. K.T. A.C.H. Y.W. T.M. and I.J.F. are co-founders and have a financial interest in Von Baer Wolff, Inc. a company focused on biofabrication of autologous human hepatocytes from stem cells technology and programming liver failure and their interests are managed by the Conflict of Interest Office at the University of Pittsburgh in accordance with their policies.
Funding Information:
We thank Stephen C. Strom for sharing human iPSCs that have been used in part of this report. We also thank the Center for Biologic Imaging at the University of Pittsburgh for technical support on the transmission electro microscopy study. This project used the UPMC Hillman Cancer Center and Tissue and Research Pathology/Pitt Biospecimen Core shared resource, which is supported in part by award P30CA047904. This project was also partially supported by the Pilot and Feasibility program under the NIH/NIDDK Digestive Disease Research Core Center grant P30DK120531 . This work was supported by NIH grants DK099257 , DK117881 , DK119973 , and TR002383 to A.S.-G., P01DK096990 to I.J.F. and A.S.G., DK107697 to E.M.S. and A.S.G., and DK103645 to A.W.D., and by the American Liver Foundation to K.T.
Publisher Copyright:
© 2020 The Author(s)
PY - 2020/6/2
Y1 - 2020/6/2
N2 - The availability of an autologous transplantable auxiliary liver would dramatically affect the treatment of liver disease. Assembly and function in vivo of a bioengineered human liver derived from induced pluripotent stem cells (iPSCs) has not been previously described. By improving methods for liver decellularization, recellularization, and differentiation of different liver cellular lineages of human iPSCs in an organ-like environment, we generated functional engineered human mini livers and performed transplantation in a rat model. Whereas previous studies recellularized liver scaffolds largely with rodent hepatocytes, we repopulated not only the parenchyma with human iPSC-hepatocytes but also the vascular system with human iPS-endothelial cells, and the bile duct network with human iPSC-biliary epithelial cells. The regenerated human iPSC-derived mini liver containing multiple cell types was tested in vivo and remained functional for 4 days after auxiliary liver transplantation in immunocompromised, engineered (IL2rg−/−) rats.
AB - The availability of an autologous transplantable auxiliary liver would dramatically affect the treatment of liver disease. Assembly and function in vivo of a bioengineered human liver derived from induced pluripotent stem cells (iPSCs) has not been previously described. By improving methods for liver decellularization, recellularization, and differentiation of different liver cellular lineages of human iPSCs in an organ-like environment, we generated functional engineered human mini livers and performed transplantation in a rat model. Whereas previous studies recellularized liver scaffolds largely with rodent hepatocytes, we repopulated not only the parenchyma with human iPSC-hepatocytes but also the vascular system with human iPS-endothelial cells, and the bile duct network with human iPSC-biliary epithelial cells. The regenerated human iPSC-derived mini liver containing multiple cell types was tested in vivo and remained functional for 4 days after auxiliary liver transplantation in immunocompromised, engineered (IL2rg−/−) rats.
KW - bioengineered human liver
KW - human iPS cells
KW - human iPS-biliary cells
KW - human iPS-endothelial cells
KW - human iPS-hepatocytes
KW - liver maturation
KW - mini human liver
KW - organ-microenvironment
KW - transplantation
UR - http://www.scopus.com/inward/record.url?scp=85085547431&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85085547431&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2020.107711
DO - 10.1016/j.celrep.2020.107711
M3 - Article
C2 - 32492423
AN - SCOPUS:85085547431
VL - 31
JO - Cell Reports
JF - Cell Reports
SN - 2211-1247
IS - 9
M1 - 107711
ER -
