Abstract
BackgroundAcute liver failure (ALF) is a life-threatening disease with a high mortality rate. However, there are limited treatments or devices available for ALF therapy. Here, we aimed to develop a new strategy for ALF treatment by transplanting functional liver organoids (LOs) generated from single donor-derived human induced pluripotent stem cell (hiPSC) endoderm, endothelial cells (ECs), and mesenchymal cells (MCs).MethodsFirst, we isolated ECs and MCs from a single donor umbilical cord (UC) through enzyme digestion and characterized the UC-ECs and UC-MCs by flow cytometry. Second, using a nonviral reprogramming method, we generated same donor-derived hiPSCs from the UC-ECs and investigated their hepatic differentiation abilities. Finally, we simultaneously plated EC-hiPSC endoderm, UC-ECs, and UC-MCs in a three-dimensional (3D) microwell culture system, and generated single donor cell-derived differentiated LOs for ALF mouse treatment.ResultsWe obtained ECs and MCs from a single donor UC with high purity, and these cells provided a multicellular microenvironment that promoted LO differentiation. hiPSCs from the same donor were generated from UC-ECs, and the resultant EC-hiPSCs could be differentiated efficiently into pure definitive endoderm and further into hepatic lineages. Simultaneous plating of EC-hiPSC endoderm, UC-ECs, and UC-MCs in the 3D microwell system generated single donor cell-derived LOs (SDC-LOs) that could be differentiated into functional LOs with enhanced hepatic capacity as compared to that of EC-hiPSC-derived hepatic-like cells. When these functional SDC-LOs were transplanted into the renal subcapsules of ALF mice, they rapidly assumed hepatic functions and improved the survival rate of ALF mice.ConclusionThese results demonstrate that functional LOs generated from single donor cells can improve the condition of ALF mice. Functional SDC-LO transplantation provides a promising novel approach for ALF therapy.
Highlights
Acute liver failure (ALF) is a life-threatening disease with a high mortality rate
Nie et al Stem Cell Research & Therapy (2018) 9:5 donor shortage [6]. This shortage in primary human hepatocyte (PHH) might be resolved with the emergence of human induced pluripotent stem cells, which could provide an unlimited source of hepatic-like cells (HLCs) [7, 8]. hiPSC-Hepatic-like cell (HLC) have been used for engraftment in a liver injury model and to repair injured liver tissue [9]; following engraftment, it took weeks for hiPSC-HLCs to start producing detectable levels of specific proteins and exhibit hepatic function [10]
Flow cytometry analysis revealed that umbilical cord (UC)-Endothelial cell (EC) and control ECs (con-ECs) displayed similar surface markers: CD31+, CD144+, CD146+, CD90−, CD45−, and SSEA4−
Summary
Acute liver failure (ALF) is a life-threatening disease with a high mortality rate. there are limited treatments or devices available for ALF therapy. As an alternative to liver transplantation, primary human hepatocyte (PHH) transplantation has been performed in patients with liver-based metabolic disease and ALF, and in neonates and children with metabolic disorders [5]. Nie et al Stem Cell Research & Therapy (2018) 9:5 donor shortage [6] This shortage in PHHs might be resolved with the emergence of human induced pluripotent stem cells (hiPSCs), which could provide an unlimited source of hepatic-like cells (HLCs) [7, 8]. Transplants that can assume hepatic functions with little delay are required for ALF treatment
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
