Gastric Fundic Glands Isolation: A Method to Isolate Fundic Glands from Human Stomach Tissue

Abstract

Source: Teal, E. et al. Mouse- and Human-derived Primary Gastric Epithelial Monolayer Culture for the Study of Regeneration. J. Vis. Exp. (2018)

This video describes a method to isolate gastric epithelial cells from the human fundic tissue. The isolated epithelial cells can be used as an in vitro model to study their regulatory roles in gastric physiology and pathophysiology.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national and international guidelines for human welfare and have been reviewed by the local institutional review board.

1. Establishing Human-derived 3D Gastric Fundic Organoids

NOTE: To avoid contamination, perform protocol in its entirety within a sterile tissue culture hood. Human fundic tissue was collected from patients undergoing sleeve gastrectomy.

1. Collect the tissue from patients undergoing sleeve gastrectomy and place it in the ice-cold Ca²⁺/Mg

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Ca2+/Mg2+ -free DPBS	Fisher	21-031CV
Sterile gauze
Curved hemostatic forceps
Sterile Petri dish
Sterile razor blades
Advanced Dulbecco's Modified Eagle/F12 medium (basal media)	Life Technologies	12634-010
Penicillin/Streptomycin	Thermo Scientific	SV30010
Amphotericin B/Gentamicin	Thermo Scientific	R01510
GlutaMAX (L-glutamine)	Life Technologies	25030-081
Kanamycin	Thermo Fisher	RO1510
Collagenase Type 1	Worthington	LS004214
Bovine Serum Albumin	Sigma Aldrich	A9418
HEPES buffer	Sigma Aldrich	H0887
125 mL Round bottom flask
1 in (25 mm) Stir bar
Rubber Septa
20G Spinal needles of 3.5 in length	Thermo Fisher	405182
Kimwipes (tissue paper)	Fischer Scientific	06-666C
Oxygen Tank with rubber hosing
Ring Stand Clamps
37 °C Water bath
5 mL Cell culture test tubes	Fisher	14-956-3C