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…
Divulgaciones
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 |
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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).