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LIVER Publication-derived

Sepsis Liver Organoids

Full SOP with Academic or a plan

Source Li et al., 2024 · The University of Tokyo, Institute of Medical Science · 10.1016/j.celrep.2024.113918

Authors: Yang Li, Yunzhong Nie, Xia Yang, Yang Liu, Xiaoshan Deng, Yoshihito Hayashi, Riana Plummer, Qinglin Li, Na Luo, Toshiharu Kasai, Takashi Okumura, Yu Kamishibahara, Takemasa Komoto, Takuya Ohkuma, Satoshi Okamoto, Yumiko Isobe, Kiyoshi Yamaguchi, Yoichi Furukawa, Hideki Taniguchi Duration: 19 days 6 phases Cell source: Human iPSC

Abstract

This protocol describes the generation of liver organoids containing Kupffer cells (KuLOs) by co-culturing human iPSC-derived erythro-myeloid progenitors (EMPs) with hiPSC-derived liver organoids. KuLOs recapitulate fetal liver hematopoiesis and can model sepsis-associated liver dysfunction when exposed to LPS and interferon-γ.

Cell source
Human iPSC
Application
Disease modeling

Protocol overview

21 steps across 6 phases

Phase 1 · Generation of EMPs from hiPSCs Days 0-8
  1. a hiPSC seeding and colony formation
  2. b Mesoderm specification
  3. c Hematopoietic specification
  4. d Collection of hiPSC-EMPs
Phase 2 · Generation of liver organoids (LOs) from hiPSCs Days 0-6
  1. a Preparation of hiPSC-derived cell components
  2. b Assembly of liver organoids in 3D microplates
  3. c Daily medium changes
Phase 3 · Co-culture of EMPs with liver organoids to generate KuLOs Days 0-14
  1. a Assembly of EMP and LO co-culture
  2. b Medium change and growth factor optimization (Days 1-6)
  3. c Transition to M-CSF supplementation (Day 6-14)
  4. d Kupffer cell characterization and KuLO maturation
Phase 4 · Modeling sepsis-associated liver dysfunction Days 14-16
  1. a Preparation of stimulation medium
  2. b Exposure of KuLOs to LPS/IFN-γ
  3. c Assessment of sepsis-like phenotype
Phase 5 · Recovery of sepsis-induced KuLO dysfunction Days 16-19
  1. a Removal of endotoxin stimulation
  2. b Daily medium changes during recovery
  3. c Assessment of functional recovery
Phase 6 · Drug screening: TAK-242 treatment during sepsis Days 14-19
  1. a Preparation of TAK-242-supplemented medium
  2. b Early TAK-242 treatment (concurrent with LPS/IFN-γ)
  3. c Assessment of TAK-242 efficacy
  4. d Accelerated recovery with TAK-242

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Attribution

This SOP was written by Organthis based on the published method in Li et al., 2024. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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