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

Published: April 30, 2023

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 undergoin…

Açıklamalar

The authors have nothing to disclose.

Materials

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

Etiketler

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Gastric Fundic Glands Isolation: A Method to Isolate Fundic Glands from Human Stomach Tissue. J. Vis. Exp. (Pending Publication), e20426, doi: (2023).

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