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LIVER Publication-derived

Liver Organoids for Drug Screening in Suspension

Full SOP with Academic or a plan

Source Harrison et al., 2023 · University of Oslo, Institute of Basic Medical Sciences, Hybrid Technology Hub-Centre of Excellence · 10.1038/s12276-023-01074-1

Authors: Sean P. Harrison, Richard Siller, Yoshiaki Tanaka, Maria Eugenia Chollet, Gareth J. Sullivan Duration: 20 days 6 phases Cell source: Human PSC (H1 ESC, 207 ESC, AG27 iPSC)

Abstract

This protocol describes a suspension-based, ECM-independent method for generating large quantities of vascularized liver organoids from human pluripotent stem cells using small-molecule patterning. The resulting organoids exhibit complex cellular repertoires including hepatocytes, cholangiocytes, endothelial cells, Kupffer cells, and stellate cells, with de novo vasculature and functional liver properties including drug metabolism, serum protein production, and coagulation factor synthesis.

Cell source
Human PSC (H1 ESC, 207 ESC, AG27 iPSC)
Application
Developmental study, disease modeling, drug screening, tissue engineering

Protocol overview

14 steps across 6 phases

Phase 1 · hPSC Culture and Preparation Maintenance
  1. a Maintain hPSC lines in feeder-free conditions
  2. b Harvest cells with Accutase
Phase 2 · Aggregate Formation (Day 0) Day 0
  1. a Seed cells for aggregate formation
  2. b Allow cells to self-organize into aggregates
Phase 3 · Mesendoderm Induction (Days 1-2) Day 0-2
  1. a Prepare aggregates for differentiation
  2. b Initiate differentiation with CHIR99021
  3. c Remove CHIR99021 and continue differentiation
Phase 4 · Hepatic Specification (Days 2-7) Day 2-7
  1. a Switch to hepatic specification medium
  2. b Culture with medium changes
Phase 5 · Liver Organoid Maturation (Days 7-20) Day 7-20
  1. a Prepare maturation medium
  2. b Resuspend aggregates in maturation medium
  3. c Perform medium exchanges
Phase 6 · Long-term Culture and Functional Assessment (Days 20+) Day 20 onwards
  1. a Continue maturation medium culture
  2. b Harvest organoids for analysis

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Attribution

This SOP was written by Organthis based on the published method in Harrison et al., 2023. Once the originating laboratory has confirmed it, the SOP is marked Verified.

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