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LIVER Publication-derived

Human iPSC-Derived Hepatic Progenitors Bioengineered into Liver Organoids using Inverted Colloidal Crystal Poly(Ethylene Glycol) Scaffold

Source Ng et al., 2018 · King's College London, Centre for Stem Cells and Regenerative Medicine & Institute for Liver Studies · 10.1016/j.biomaterials.2018.07.043

👤 Soon Seng Ng, Kourosh Saeb-Parsy, Samuel J.I. Blackford, Joe M. Segal, Maria Paola Serra, Marta Horcas-Lopez, Da Yoon No, Sotiris Mastoridis, Wayel Jassem, Curtis W. Frank, Nam Joon Cho, Hiromitsu Nakauchi, Jeffrey S. Glenn, S. Tamir Rashid ⏱ 14 days 📋 3 phases 🧫 Human iPSC-derived hepatic progenitors

Abstract

This protocol describes the generation of liver organoids from human iPSC-derived hepatic progenitors cultured in inverted colloidal crystal (ICC) scaffolds made from polyethylene glycol. The organoids exhibit advanced hepatic function, morphology, and gene expression profiles comparable to fetal liver tissue, with potential applications in drug metabolism studies, disease modeling, and transplantation.

Cell source
Human iPSC-derived hepatic progenitors
Application
Liver organoid generation and disease modeling

Protocol overview

21 steps across 3 phases

ICC Scaffold Fabrication Day 0 (Preparation)
  1. 1 Construct sacrificial crystal lattice
  2. 2 Anneal polystyrene lattice
  3. 3 Prepare PEGDA precursor solution
  4. 4 Infiltrate lattice with PEGDA precursor
  5. 5 Perform free-radical polymerization
  6. 6 Remove sacrificial polystyrene lattice
  7. 7 Equilibrate ICC scaffolds in deionized water
  8. 8 Coat ICC scaffolds with ECM proteins
  9. 9 Prepare tubular ICC scaffolds sized for use in 96-well plate format.
iPSC-Derived Hepatic Progenitor (IH) Cell Seeding and Initial Attachment (Phase I of Organogenesis) Day 1–5
  1. 1 Generate iPSC-derived hepatic progenitors (IH)
  2. 2 Prepare cell suspension at target density
  3. 3 Seed cells onto ICC scaffolds
  4. 4 Incubate for initial cell attachment (dry seeding phase)
  5. 5 Transfer scaffolds to culture plate and refresh media
  6. 6 Refresh media at 4 hours post-seeding
  7. 7 Monitor Phase I morphology (Days 0–5)
Organoid Formation and Maturation (Phase II of Organogenesis) Day 7–14
  1. 1 Continue culture and monitor organoid formation
  2. 2 Refresh media every two days
  3. 3 Monitor Phase II morphology and organoid maturation (Days 7–14)
  4. 4 Assess hepatic function and maturation markers
  5. 5 Optional: Harvest organoids for downstream analyses

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Ng et al., 2018. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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