Skip to content
← Back to browse
LIVER Publication-derived

Scalable production of tissue-like vascularized liver organoids from human PSCs

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

👤 Sean P. Harrison, Richard Siller, Yoshiaki Tanaka, Maria Eugenia Chollet, Gareth J. Sullivan ⏱ 20 days 📋 6 phases 🧫 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

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

Full SOP

🔬

Create a free account to access this protocol

Join OrganMatch to unlock step-by-step procedures, reagent concentrations, QC checklists, and downloadable batch record templates.

Create free account

Already registered? Log in

Attribution

This SOP was authored by Organthis based on the published method in Harrison et al., 2023. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

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