Here, a method using a permeable membrane insert-based infection system to study the effects of Streptolysin S, a secreted toxin produced by Group A Streptococcus, on keratinocytes is described. This system can be readily applied to the study of other secreted bacterial proteins on various host cell types during infection.
Total time
~3-4 days (including bacterial culture, infection, and sample collection)
Model organism
Keratinocytes (mammalian host cells); Group A Streptococcus
Steps
1
Culture Group A Streptococcus bacteria
Prepare and grow Group A Streptococcus cultures under appropriate conditions prior to infection experiments.
▶ 00:58
2
Infect host cells using permeable membrane insert system
Establish keratinocyte cultures in permeable membrane inserts and expose them to Group A Streptococcus to study secreted toxin effects in a controlled compartmentalized setup.
▶ 01:48
3
Collect samples from infected cultures
Harvest cells and media from both sides of the membrane insert system for downstream analysis of toxin-induced cellular changes.
▶ 03:52
4
Analyze effects of secreted bacterial toxins on host cells
Examine collected samples to quantify and characterize the impact of Streptolysin S and other secreted Group A Streptococcus proteins on keratinocyte integrity and function.
▶ 06:13