Assembly and Function of a Bioengineered Human Liver for Transplantation Generated Solely from Induced Pluripotent Stem Cells

Kazuki Takeishi; Alexandra Collin de l'Hortet; Yang Wang; Kan Handa; Jorge Guzman-Lepe; Kentaro Matsubara; Kazutoyo Morita; Sae Jang; Nils Haep; Rodrigo M Florentino; Fangchao Yuan; Ken Fukumitsu; Kimimasa Tobita; Wendell Sun; Jonathan Franks; Evan R Delgado; Erik M Shapiro; Nicolas A Fraunhoffer; Andrew W Duncan; Hiroshi Yagi; Tomoji Mashimo; Ira J Fox; Alejandro Soto-Gutierrez

doi:10.1016/j.celrep.2020.107711

Assembly and Function of a Bioengineered Human Liver for Transplantation Generated Solely from Induced Pluripotent Stem Cells

Cell Rep. 2020 Jun 2;31(9):107711. doi: 10.1016/j.celrep.2020.107711.

Authors

Kazuki Takeishi¹, Alexandra Collin de l'Hortet², Yang Wang³, Kan Handa², Jorge Guzman-Lepe², Kentaro Matsubara², Kazutoyo Morita², Sae Jang², Nils Haep², Rodrigo M Florentino⁴, Fangchao Yuan⁵, Ken Fukumitsu², Kimimasa Tobita⁶, Wendell Sun⁷, Jonathan Franks⁸, Evan R Delgado⁹, Erik M Shapiro¹⁰, Nicolas A Fraunhoffer¹¹, Andrew W Duncan⁹, Hiroshi Yagi¹², Tomoji Mashimo¹³, Ira J Fox¹⁴, Alejandro Soto-Gutierrez¹⁵

Affiliations

¹ Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Surgery and Science, Graduate School of Medical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
² Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA.
³ Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Hepatobiliary Surgery, Peking University People's Hospital, Beijing 100044, China.
⁴ Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Physiology and Biophysics, Universidade Federal de Minas Gerais, Belo Horizonte 31270-010, Brazil.
⁵ Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China.
⁶ Department of Bioengineering and Department of Developmental Biology, University of Pittsburgh, Pittsburgh, PA 15201, USA.
⁷ LifeCell Corporation, Branchburg, NJ 08876, USA.
⁸ Center for Biologic Imaging, University of Pittsburgh Medical School, Pittsburgh, PA 15261, USA.
⁹ Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219-3110, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, USA.
¹⁰ Department of Radiology, Michigan State University, East Lansing, MI 48824, USA.
¹¹ Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; Facultad de Ciencias de la Salud, Carrera de Medicina, Universidad Maimónides, Ciudad Autónoma de Buenos Aires and Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Ciudad Autónoma de Buenos Aires, Buenos Aires 1001, Argentina.
¹² Department of Surgery, School of Medicine, Keio University, Tokyo 160-8582, Japan.
¹³ Division of Animal Genetics, Laboratory Animal Research Center, Institute of Medical Science, University of Tokyo, Tokyo 158-8557, Japan.
¹⁴ McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219-3110, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, USA; Department of Surgery, Children's Hospital of Pittsburgh of UPMC, University of Pittsburgh, Pittsburgh, PA 15224, USA.
¹⁵ Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, USA; McGowan Institute for Regenerative Medicine, University of Pittsburgh, Pittsburgh, PA 15219-3110, USA; Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, USA. Electronic address: als208@pitt.edu.

Abstract

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.

Keywords: bioengineered human liver; human iPS cells; human iPS-biliary cells; human iPS-endothelial cells; human iPS-hepatocytes; liver maturation; mini human liver; organ-microenvironment; transplantation.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Activins / genetics
Activins / metabolism
Animals
Bone Morphogenetic Protein 4 / genetics
Bone Morphogenetic Protein 4 / metabolism
Cell Differentiation
Cells, Cultured
Cellular Reprogramming
Fetus / cytology
Fibroblast Growth Factor 2 / genetics
Fibroblast Growth Factor 2 / metabolism
Fibroblasts / cytology
Fibroblasts / metabolism
Hepatocytes / cytology
Hepatocytes / metabolism
Hepatocytes / transplantation*
Humans
Immunocompromised Host
Induced Pluripotent Stem Cells / cytology
Induced Pluripotent Stem Cells / metabolism
Male
Rats
Rats, Sprague-Dawley
Tissue Engineering*
Tissue Scaffolds / chemistry
Transcription Factors / genetics
Transcription Factors / metabolism

Substances

Bone Morphogenetic Protein 4
Transcription Factors
activin A
Fibroblast Growth Factor 2
Activins

Abstract

Publication types

MeSH terms

Substances

Grants and funding