Skip to content
Back to browse
GASTRIC Publication-derived

Gastric Organoids for Disease Modelling

Full SOP with Academic or a plan

Source Mccracken et al., 2017 · Cincinnati Children's Hospital Medical Center · 10.1038/nature21021

Authors: Kyle W. Mccracken, Eitaro Aihara, Baptiste Martin, Calyn M. Crawford, Taylor Broda, Julie Treguier, Xinghao Zhang, John M. Shannon, Marshall H. Montrose, James M. Wells Duration: 34 days 5 phases Cell source: Human iPSC (H1 hESC-derived)

Abstract

This protocol describes the directed differentiation of human pluripotent stem cells (hPSCs) into three-dimensional fundic gastric organoids (hFGOs) containing fundus-like epithelium with parietal, chief, and mucous cells. By recapitulating Wnt/β-catenin signaling pathways identified in embryonic stomach development, the method enables generation of functional human fundus tissue for studying gastric physiology and disease.

Cell source
Human iPSC (H1 hESC-derived)
Application
Disease modeling; developmental study of fundic epithelium

Protocol overview

9 steps across 5 phases

Phase 1 · Definitive Endoderm (DE) Differentiation Day 0-3
  1. a Dissociate hPSCs into single cells
  2. b Induce definitive endoderm with Activin A
Phase 2 · Posterior Foregut Specification Day 3-6
  1. a Differentiate DE to posterior foregut with CHIR, FGF4, and Noggin
Phase 3 · Three-Dimensional Culture and Gastric Specification (Fundus Lineage) Day 6-20
  1. a Transition to three-dimensional culture in Matrigel
  2. b Specify fundus identity with CHIR, Noggin, and RA (Days 6-9)
  3. c Continue fundus specification with CHIR and FGF10 (Days 9-20)
Phase 4 · Organoid Growth and Maintenance (Days 20-30) Day 20-30
  1. a Maintain hFGOs with CHIR and FGF10
Phase 5 · Parietal Cell Induction (Days 30-34) Day 30-34
  1. a Screen or apply parietal cell-inducing conditions (Optional)
  2. b Continue basal culture to day 34

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 Mccracken et al., 2017. 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