Examination of Pyroptosis by Flow Cytometry

The objective of our research team is to determine whether avian cells undergo pyroptosis similar to that of mammalian cells. Furthermore, we aim to determine pyroptosis through more accurate and easier means. Many genes in avian species significantly differ from those in mammals, making it difficult to use reagents commonly employed in mammalian cell studies for avian cell death research.

Thus, the research on avian cell death is much more difficult than that on human or murine cells. We utilize the characteristic plasma membrane pores in pyroptotic cells formed by gasdermin N-terminal fragments to develop a flow cytometric-based strategy for the determination of pyroptosis. By this novel approach, we observed the occurrence of infectious-bursal-disease-induced pyroptosis in avian cells.

In contrast to previous methods such as western blot or AFM analysis, our protocol is able to accurately quantify the percentage of pyroptotic cells by two parameters concurrently, the plasma membrane pore formation and the plasma membrane rupture, facilitating to differentiate cells with pyroptosis from other forms of cell death.