Skip to content
Back to browse
LIVER Publication-derived

Liver Organoids for Drug Screening (2017)

Full SOP with Academic or a plan

Source Sgodda et al., 2017 · Hannover Medical School · 10.1089/scd.2017.0023

Authors: Malte Sgodda, Zhen Dai, Robert Zweigerdt, Amar Deep Sharma, Michael Ott, Tobias Cantz Duration: 21 days 5 phases Cell source: Human ESC (H9 line) and Human iPSC

Abstract

This protocol describes a scalable 3D suspension culture system for generating hepatic organoids from pluripotent stem cell-derived hepatic progenitors. The method maintains cells for up to three weeks with stable metabolic features and enables sensitive detection of acetaminophen-related hepatotoxicity, a common source of drug-induced liver failure.

Cell source
Human ESC (H9 line) and Human iPSC
Application
Disease modeling, Drug screening, Hepatotoxicity assessment

Protocol overview

19 steps across 5 phases

Phase 1 · Maintenance and Initial Differentiation of PSCs Days 0-3
  1. a Prepare hESC culture substrate
  2. b Passage and prepare monolayer for differentiation
  3. c Add CHIR99021 for Wnt signaling modulation
  4. d Continue cultivation for three days
Phase 2 · Hepatic Progenitor Differentiation Days 4-12
  1. a First hepatic differentiation medium change
  2. b Continue with daily medium changes
  3. c Second hepatic differentiation phase with HGF
  4. d Final maturation with HGF, OncostatinM, and Dexamethasone
Phase 3 · Organoid Formation and Suspension Culture Days 16 onwards
  1. a Mechanical dissection of monolayer
  2. b Transfer to suspension culture
  3. c Monitor organoid self-assembly
  4. d Size control and separation at 48 hours
  5. e Return sorted organoids to suspension culture
Phase 4 · Improved Medium Formulation for Enhanced Hepatic Characteristics Days 16 onwards (optional optimization)
  1. a Prepare modified hepatocyte culture medium
  2. b Culture organoids in modified medium
Phase 5 · Hepatotoxicity Assessment with Acetaminophen Day 21 (or after three weeks culture)
  1. a Prepare organoids for toxicity assay
  2. b Plate organoids in 24-well plates
  3. c Add acetaminophen at indicated concentrations
  4. d Perform WST-1 cell viability assay

Full SOP

Full SOPs need an account and a plan

Create an account, then get free Academic access (for verified academics) or choose a plan. Plans include step-by-step procedures, reagent concentrations, QC checklists and batch record templates.

Create an account

Already registered? Log in

Attribution

This SOP was written by Organthis based on the published method in Sgodda et al., 2017. Once the originating laboratory has confirmed it, the SOP is marked Verified.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence