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Original research
Three-dimensional bioprinted hepatorganoids prolong survival of mice with liver failure
  1. Huayu Yang1,
  2. Lejia Sun1,
  3. Yuan Pang2,3,4,
  4. Dandan Hu1,5,
  5. Haifeng Xu1,
  6. Shuangshuang Mao2,3,4,
  7. Wenbo Peng6,
  8. Yanan Wang7,
  9. Yiyao Xu1,
  10. Yong-Chang Zheng1,
  11. Shunda Du1,
  12. Haitao Zhao1,
  13. Tianyi Chi1,
  14. Xin Lu1,
  15. Xinting Sang1,
  16. Shouxian Zhong1,
  17. Xin Wang8,9,10,
  18. Hongbing Zhang7,
  19. Pengyu Huang6,11,
  20. Wei Sun2,3,4,12,
  21. Yilei Mao1
  1. 1Department of Liver Surgery, Peking Union Medical College (PUMC) Hospital, PUMC & Chinese Academy of Medical Sciences, Beijing, China
  2. 2Biomanufacturing Center, Department of Mechanical Engineering, Tsinghua University, Beijing, China
  3. 3Biomanufacturing and Rapid Forming Technology Key Laboratory of Beijing, Tsinghua University, Beijing, China
  4. 4Overseas Expertise Introduction Center for Discipline Innovation, Tsinghua University, Beijing, China
  5. 5Department of Hepatobiliary Surgery, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong, China
  6. 6School of Life Science and Technology, ShanghaiTech University, Shanghai, China
  7. 7State Key Laboratory of Medical Molecular Biology, Department of Physiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, Beijing, China
  8. 8Research Center for Laboratory Animal Science, Inner Mongolia University, Hohhot, Inner Mongolia, China
  9. 9Hepatoscience Section, Cell Lab Tech Inc, Sunnyvale, California, USA
  10. 10Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis, Minnesota, USA
  11. 11Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, Shanghai, China
  12. 12Department of Mechanical Engineering, Drexel University, Philadelphia, Pennsylvania, USA
  1. Correspondence to Professor Yilei Mao, Department of Liver Surgery, Peking Union Medical College (PUMC) Hospital, PUMC & Chinese Academy of Medical Sciences, Beijing, China; pumch-liver{at}hotmail.com; Professor Wei Sun, Biomanufacturing Center, Department of Mechanical Engineering, Tsinghua University, Beijing, China; weisun{at}tsinghua.edu.cn; Professor Pengyu Huang, School of Life Science and Technology, ShanghaiTech University, Shanghai, China; huangpy{at}shanghaitech.edu.cn

Abstract

Objective Shortage of organ donors, a critical challenge for treatment of end-stage organ failure, has motivated the development of alternative strategies to generate organs in vitro. Here, we aim to describe the hepatorganoids, which is a liver tissue model generated by three-dimensional (3D) bioprinting of HepaRG cells and investigate its liver functions in vitro and in vivo.

Design 3D bioprinted hepatorganoids (3DP-HOs) were constructed using HepaRG cells and bioink, according to specific 3D printing procedures. Liver functions of 3DP-HOs were detected after 7 days of differentiation in vitro, which were later transplanted into Fah-deficient mice. The in vivo liver functions of 3DP-HOs were evaluated by survival time and liver damage of mice, human liver function markers and human-specific debrisoquine metabolite production.

Results 3DP-HOs broadly acquired liver functions, such as ALBUMIN secretion, drug metabolism and glycogen storage after 7 days of differentiation. After transplantation into abdominal cavity of Fah-/-Rag2-/- mouse model of liver injury, 3DP-HOs further matured and displayed increased synthesis of liver-specific proteins. Particularly, the mice acquired human-specific drug metabolism activities. Functional vascular systems were also formed in transplanted 3DP-HOs, further enhancing the material transport and liver functions of 3DP-HOs. Most importantly, transplantation of 3DP-HOs significantly improved the survival of mice.

Conclusions Our results demonstrated a comprehensive proof of principle, which indicated that 3DP-HO model of liver tissues possessed in vivo hepatic functions and alleviated liver failure after transplantation, suggesting that 3D bioprinting could be used to generate human liver tissues as the alternative transplantation donors for treatment of liver diseases.

  • liver failure
  • liver transplantation
  • liver function test
  • liver metabolism
  • liver regeneration
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Footnotes

  • HY, LS, YP and DH contributed equally.

  • Contributors YM, WS and PH conceived the project. HY, LS, YP and DH performed most of the experiments. YP and SM optimised the printing process of 3DP-HOs. YW, YX, YZ and SD analysed the in vitro functions of 3DP-HOs. HZhao and TC analysed gene expression of 3DP-HOs. XL, XS, SZ, XW and HZhang designed the experiments to characterize in vivo functions of 3DP-HOs. HY, LS, HX and WP performed the in vivo experiments. HY, LS, YM, WS and PH analysed the data. HY, PH, HZhang and YP wrote the manuscript.

  • Funding This work was supported by grants from the National High-tech Research and Development Projects (863) (no.2015AA020303), National Basic Research Program of China 973 Program Grants (2015CB553802) and CAMS Innovation Fund for Medical Sciences (CIFMS) (No.2016-I2M-1-001). This work was also supported by grants from Strategic Priority Program (SPP) on Space Science (no. XDA15014300), 111 Project (no. B17026), the Ministry of Science and Technology of China (MOST; 2019YFA0801501, 2016YFA0100500), the National Natural Science Foundation of China (NSFC; 31970687, 31571509, 31522038, 51805294, 81730078), ShanghaiTech start-up grant to Pengyu Huang and International Science and Technology Projects (2016YFE0107100).

  • Competing interests The 3D bioprinter was provided by HEALTH Biomed Company.

  • Patient and public involvement Patients and/or the public were not involved in the design, or conduct, or reporting, or dissemination plans of this research.

  • Patient consent for publication Not required.

  • Provenance and peer review Not commissioned; externally peer reviewed.

  • Data availability statement All data relevant to the study are included in the article or uploaded as online supplementary information. Other data are available on reasonable request.

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