An adapted novel flow cytometry methodology to delineate types of cell death in airway epithelial cells
2Telethon Kids Institute, University of Western Australia, Western Australia 6009, Australia
3Department of Respiratory and Sleep Medicine, Perth Children’s Hospital, Western Australia 6009, Australia
4School of Public Health, Curtin University, Western Australia 6102, Australia
5Centre for Cell Therapy and Regenerative Medicine, School of Medicine and Pharmacology, University of Western Australia, Western Australia 6009, Australia
INTRODUCTION
MATERIAL AND METHODS
Study population
Cell culture
Rhinovirus infection
Flow cytometry
Gating strategies
Statistics
RESULTS
A5/PI staining cannot sufficiently differentiate between cell populations using flow cytometry
A5/TP3 staining allows differentiation between six cell populations following fixation
A5/TP3 staining allowed measurement of viable, necrotic, and apoptotic events in primary AEC after fixation and RV infection
A5/TP3 staining allows the observation in phenotypical differences in apoptotic cell disassembly
Populations | Annexin V | TO-PRO-3 | FSC/SSC |
---|---|---|---|
Viable | Low | Low | Intermediate/high |
Necrotic | High | High | Intermediate/high |
A5+ apoptotic | High | Intermediate | Intermediate/high |
A5− apoptotic | Low | Intermediate | Intermediate/high |
Apoptotic bodies | Intermediate | Low/intermediate | Low/low |
Debris | Low | Low | Low |
DISCUSSION
Acknowledgments
References
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