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LIVER Publication-derived

A Scalable Approach for the Generation of Human Pluripotent Stem Cell-Derived Hepatic Organoids with Sensitive Hepatotoxicity Features

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

👤 Malte Sgodda, Zhen Dai, Robert Zweigerdt, Amar Deep Sharma, Michael Ott, Tobias Cantz ⏱ 21 days 📋 5 phases 🧫 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

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

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Sgodda et al., 2017. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

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