Three-dimensional bioprinted hepatorganoids prolong survival of mice with liver failure

Abstract

ObjectiveShortage 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.Design3D 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.Results3DP-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.ConclusionsOur 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.

3D Bioprinting of Vascularized Tissues for in vitro and in vivo Applications

“…Recently, Yang et al (2020) successfully printed functional liver tissue that exhibited normal liver functions such as ALBUMIN secretion, drug metabolism, and glycogen storage after 7 days of cell differentiation and growth in mouse models. Although no vascular designs were incorporated into the tissue constructs, the printing material used (hepatorganoids with the combination of HepaRG cells and bioink), has the potential to result in the development of functional vascular networks in transplanted tissues ( Yang et al, 2020 ). The growth of the vasculature improved nutrient support and liver functions in the 3D printed construct , ultimately leading to the prolonged survival of mice with liver failure ( Yang et al, 2020 ).…”

“…Although no vascular designs were incorporated into the tissue constructs, the printing material used (hepatorganoids with the combination of HepaRG cells and bioink), has the potential to result in the development of functional vascular networks in transplanted tissues ( Yang et al, 2020 ). The growth of the vasculature improved nutrient support and liver functions in the 3D printed construct , ultimately leading to the prolonged survival of mice with liver failure ( Yang et al, 2020 ).…”

Microfluidic Organoids-on-a-Chip: Quantum Leap in Cancer Research

“…Moreover, human liver cancer organoids derived from needle biopsies recapitulated the histology and molecular profile of primary tumors [53]. Recently, Yang et al established hepatorganoids that prolong mice's survival with liver failure using 3D bio-printing technology, which also allows for performing drug screening [97]. This method was further extended to generate HBV infected patient-derived models for HCC.…”

Current Advances in 3D Bioprinting Technology and Its Applications for Tissue Engineering

“…They demonstrated that a nozzle with an inner diameter of 100 µm could be utilized without clogging. Yang et al bioprinted 3D liver functional tissues using HepaRG cells that combined gelatin and alginate composite bio-inks [ 191 ]. After verification of the 3D liver-like functional activity using the 3D bioprinted hepatoganoids, the construct was implanted into mice, and it was observed that the mice prolonged their survival in the experimental groups.…”