Gastric Organoids for Developmental Studies (2019)
Abstract
This protocol describes the preparation of optimized culture media for gastric organoid differentiation and maintenance. The media formulation incorporates basal Advanced DMEM/F12 supplemented with growth factors (R-spondin 1, Noggin, EGF), hormones (Gastrin, Prostaglandin E2), signaling modulators (A-83-01, SB202190), and supportive components (N2, B27, Glutamax, HEPES) to support sustained gastric epithelial organoid development.
Protocol overview
19 steps across 7 phases
- a Prepare Advanced DMEM/F12 base
- b Add HEPES buffer
- c Add Glutamax supplement
- d Add antibiotic protection
- a Add N2 supplement
- b Add B27 supplement
- a Prepare and add R-spondin 1
- b Prepare and add Noggin
- c Prepare and add EGF
- a Add n-Acetylcysteine
- b Add Niacinamide
- a Prepare and add A-83-01
- b Prepare and add SB202190
- a Prepare and add Gastrin
- b Prepare and add Prostaglandin E2
- a Mix media thoroughly
- b Filter-sterilize the media
- c Check media pH
- d Store the media
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Attribution
This SOP was written by Organthis based on the published method. Once the originating laboratory has confirmed it, the SOP is marked Verified.
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