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GASTRIC Publication-derived

Wnt/β-catenin promotes gastric fundus specification in mice and humans

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

👤 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 ⏱ 34 days 📋 5 phases 🧫 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

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

Full SOP

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Attribution

This SOP was authored by Organthis based on the published method in Mccracken et al., 2017. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

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