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Molecular Characterization of Gastric Epithelial Cells Using Flow Cytometry
Kevin A Bockerstett1, Chun Fung Wong2, Sherri Koehm3
1Department of Molecular Microbiology and Immunology, Saint Louis University School of Medicine, Saint Louis, MO 63104, USA. kevin.bockerstett@slu.edu.
International Journal of Molecular Sciences
|April 13, 2018
Summary
Researchers developed a new method to isolate viable single gastric epithelial cells for flow cytometry analysis. This technique aids in studying chronic gastritis and gastric cancer progression by examining cell responses to inflammation.
Area of Science:
- Gastroenterology
- Cell Biology
- Immunology
Background:
- Analyzing individual gastric epithelial cells is crucial for understanding gastric diseases like chronic gastritis and cancer.
- Previous methods faced limitations in isolating viable cells and analyzing flow cytometry data due to cell variability and autofluorescence.
Purpose of the Study:
- To develop a reliable method for isolating viable single cells from the stomach corpus.
- To optimize flow cytometry analysis for gastric epithelial cells from healthy and diseased tissues.
Main Methods:
- A two-stage enzymatic digestion using collagenase and Dispase II to disperse gastric glands into single cells.
- Flow cytometry analysis incorporating light scatter, size exclusion, doublet discrimination, viability staining, and fluorescent antibodies/lectins.
Main Results:
- Successfully isolated over 1x10^6 viable single cells per gastric corpus, representing all major cell lineages.
- Identified parietal cells and demonstrated that chronically inflamed gastric epithelial cells upregulate MHC I and II, but not CD80/CD86.
Conclusions:
- The described method enables reliable isolation and flow cytometry analysis of gastric epithelial cells.
- Chronic inflammation impacts gastric epithelial cells by upregulating MHC I and II, providing insights into gastric disease mechanisms.
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