Skip to content
Back to browse
LIVER Publication-derived

Drug-Induced Liver Injury Organ-on-Chip Models in High-Throughput Format

Full SOP with Academic or a plan

Source Zhang et al., 2023 · University of Michigan · 10.1016/j.jhep.2023.01.019

Authors: Charles J. Zhang, Sophia R. Meyer, Matthew J. O'Meara, Sha Huang, Meghan M. Capeling, Daysha Ferrer-Torres, Charlie J. Childs, Jason R. Spence, Robert J. Fontana, Jonathan Z. Sexton Duration: 14 days 10 phases Cell source: Human iPSC (fibroblast-derived)

Abstract

This protocol describes the generation and characterization of human liver organoids (HLOs) derived from iPSC lines in two complementary formats: dispersed HLOs in 384-well plates for high-throughput screening and intact HLOs adapted to a liver-on-chip system (PaDLOC) for enhanced physiological assessment. Both platforms enable identification of hepatotoxic compounds and prediction of synergistic drug toxicities through biochemical assays, morphological profiling, and single-cell transcriptomics.

Cell source
Human iPSC (fibroblast-derived)
Application
Disease modeling and drug screening for drug-induced liver injury (DILI) risk prediction

Protocol overview

35 steps across 10 phases

Phase 1 · Preparation of PaDLOC (Patient-Derived Liver-On-Chip) System Pre-seeding
  1. a Prepare Chip S1 channels with extracellular matrix
  2. b Prepare dispersed HLO cell suspension
Phase 2 · PaDLOC Cell Seeding and Attachment Day 0
  1. a Seed cells into bottom channel
  2. b Allow bottom channel cell attachment
  3. c Seed cells into top channel
  4. d Allow top channel cell attachment
  5. e Wash both channels with hepatocyte growth media
Phase 3 · PaDLOC Culture with Flow Days 1-7
  1. a Attach chip to Pod Portable Module
  2. b Place loaded pods into Zoë Culture Module
  3. c Introduce media flow at constant rate
  4. d Maintain media and culture for 7 days pre-treatment
Phase 4 · PaDLOC Drug Treatment Days 8-14
  1. a Replace media with drug-containing media
  2. b Maintain constant flow during 7-day treatment period
  3. c Collect media outflow at designated time points
  4. d Measure ALT, AST, and albumin in collected media
Phase 5 · PaDLOC Imaging and Morphological Analysis Day 14 (post-treatment)
  1. a Fix and stain PaDLOC cells
  2. b Acquire confocal microscopy images
  3. c Analyze morphological features
Phase 6 · PaDLOC Single-Cell RNA Sequencing Day 14 (post-treatment)
  1. a Harvest cells from treated PaDLOCs
  2. b Perform single-cell RNA sequencing
  3. c Analyze differential expression
Phase 7 · 384-Well Dispersed HLO Culture and Drug Screening Days 0-5
  1. a Coat 384-well plates with collagen type I
  2. b Seed dispersed HLO cells into 384-well plates
  3. c Allow cells to adhere
Phase 8 · 384-Well Drug Dispensing and Dose-Response Assessment Days 2-5
  1. a Prepare compound stocks
  2. b Dispense compounds using digital dispenser
  3. c Incubate cells with compounds
Phase 9 · 384-Well Fixation and Staining Day 5 (post-treatment)
  1. a Fix cells in 384-well plates
  2. b Permeabilize and block
  3. c Stain with fluorescent dyes and antibodies
  4. d Wash and prepare for imaging
Phase 10 · 384-Well Automated Confocal Imaging and Analysis Day 5-6 (post-staining)
  1. a Acquire confocal microscopy images
  2. b Perform image segmentation and feature extraction
  3. c Quantify cell viability and calculate IC50 values
  4. d Perform dimensionality reduction and clustering

Full SOP

Full SOPs need an account and a plan

Create an account, then get free Academic access (for verified academics) or choose a plan. Plans include step-by-step procedures, reagent concentrations, QC checklists and batch record templates.

Create an account

Already registered? Log in

Attribution

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

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