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LIVER Publication-derived

CF/PSC Model (Sampaziotis/Vallier)

Source Sampaziotis et al. · Wellcome-MRC Cambridge Stem Cell Institute / University of Cambridge · 10.1038/nbt.3275

👤 Sampaziotis F, de Brito MC, Madrigal P, Bertero A, Saeb-Parsy K, Soares FAC, Vallier L ⏱ 22 days 📋 6 phases 🧫 Human iPSC → Cholangiocytes

Abstract

Sampaziotis und Vallier (Cambridge) differenzierten iPSCs über Definitive Endoderm → Foregut → Cholangiocyten-Progenitoren zu reifen Cholangiocyten-Organoids. Die Organoids zeigten primäre Zilien, biliäre Transportproteine und vor allem CFTR-Kanalaktivität im Forskolin-Swelling-Assay. Patient-spezifische Reaktionen auf CFTR-Modulatoren wurden bei CF-Patienten unterschiedlich gerettet — direkter klinischer Nutzen für Präzisionsmedizin.

Cell source
Human iPSC → Cholangiocytes
Application
Cystic Fibrosis Drug Screening

Protocol overview

14 steps across 6 phases

hiPSC Maintenance Ongoing
  1. 1 Culture human induced pluripotent stem cells
Foregut Progenitor Generation Day 0–4 (variable)
  1. 1 Differentiate hiPSCs to Foregut Progenitors
Hepatoblast Generation Day 4–8
  1. 1 Generate hepatoblasts from foregut progenitors
Cholangiocyte Progenitor Specification Day 8–12
  1. 1 Induce biliary specification in hepatoblasts
CLC Maturation in 3D Culture Day 12–22
  1. 1 Passage cholangiocyte progenitors
  2. 2 Prepare 3D culture matrix
  3. 3 Plate cell-matrix mixture
  4. 4 Allow gel polymerization
  5. 5 Overlay with culture medium
  6. 6 Culture organoids with media changes
Optional: Signaling Pathway Inhibition Studies Day 12–22
  1. 1 Prepare three experimental conditions
  2. 2 Distribute aliquots to 24-well plates
  3. 3 Overlay with inhibitor-supplemented medium
  4. 4 Culture for 10 days and count organoids

Full SOP

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Attribution

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

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