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LIVER Publication-derived

Biliary Models for Drug Screening

Full SOP with Academic or a plan

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

Authors: Sampaziotis F, de Brito MC, Madrigal P, Bertero A, Saeb-Parsy K, Soares FAC, Vallier L Duration: 22 days 6 phases Cell source: 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

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

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Attribution

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

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