I am treating AR42J cells with 10-8 M cholecystokinin (CCK) for different time intervals up to 8 hours. Following this, I need to check for markers of cell death and for activation of the Unfolded Protein Response (UPR). I have used Annexin-V staining, Alamar Blue Viability assay and Calcein-AM staining for cell death, and not found any significant differences between the treated and control cells.
I did not find any significant changes in UPR markers as well. The markers I have checked are ATF6, GRP78, PERK, phospho-PERK, IRE1, phospho-IRE1, XBP1s, CHOP, ERO1L, OS-9 and PDI.
I am right now checking the levels of pancreatic lipase and amylase in these cells after treatment with CCK which would be the most important marker to comment on acute pancreatitis like phenotype.
I need suggestion for treatment that I should give to AR42J cells to hyper-stimulate zymogen production and secretion.
Thanks a lot for taking time to read this and for your valuable suggestions.
Best regards.
Try a dose-response (i.e. survival) curve with cholecystokinin (CCK). It could be your dose is too low. You have no evidence for cell death in any of your expts, this sugests your dose is too low. Try different doses and let the cells go 8 hrs and then check for significant cell death.
Hi Terry,
Thanks very much for your answer. Well, I missed out on informing that I have done a dose response from 10-6 to 10-8 M CCK, and time kinetics up to 8 hours. I have not seen significant or consistent change in number of dead cells in any of the condition. All higher time points are similar to 4 hour time point.
Overall, I am not sure if these cells are suitable as a cellular model for studying acute pancreatitis, specifically the role of ER stress in acute pancreatitis.
Thanks again for your response.
Best regards.
- Badges
- Science topic
- Similar topics
- Methodology
- Laboratory
- Cellular Biology Techniques