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Use of flow cytometry to quantify mouse gastric epithelial cell populations
Y Zavros1, M Van Antwerp, J L Merchant
1Howard Hughes Medical Institute, and the Department of Internal Medicine, University of Michigan, Ann Arbor 48109, USA.
Digestive Diseases and Sciences
|July 6, 2000
Summary
Mechanical dissociation and Percoll gradient centrifugation effectively isolate viable mouse gastric cells for flow cytometry. This method improves cell viability and enables precise quantification of rare cell populations.
Area of Science:
- Cell Biology
- Immunology
- Gastroenterology
Background:
- Flow cytometry requires high-quality, intact, viable cells for accurate quantification.
- Existing methods for isolating mouse gastric cells for flow cytometry are not well-established or evaluated.
Purpose of the Study:
- To develop and validate an effective method for isolating intact, viable mouse gastric cells for flow cytometry.
- To compare enzymatic separation versus mechanical dissociation for gastric cell isolation.
Main Methods:
- Mouse stomach and spleen cells were isolated using enzymatic separation or mechanical dissociation.
- Percoll density gradient centrifugation was employed to enrich viable cells and remove debris.
- Cells were analyzed by flow cytometry after labeling with fluorescent antibodies.
Main Results:
- Mechanical dissociation yielded significantly higher viable cells (10-20%) compared to enzymatic separation (1%).
- Percoll gradient centrifugation further increased viable cell percentages to 50-80%.
- Specific gastric epithelial cell populations (gastrin, somatostatin, parietal) and lymphocytes were quantified.
Conclusions:
- Mechanical dissociation followed by Percoll gradient centrifugation is the preferred method for isolating viable, intact gastric epithelial cells for flow cytometry.
- This optimized method significantly improves cell yield and purity, enabling detection of rare cell populations.