Objective of the study: Evaluate whether normal bronchial epithelial cells acquire invasiveness following exposure to particulate matter.
Initial approach: Expose the BEAS2B cell line to PM10 at concentrations of 50ug and 75ug for 48 hours. Subsequently, perform a wound healing assay and an invasion assay using matrigel.
Problem encountered: Post exposure, a significant number of cells died, resulting in inadequate cell viability. This has hindered our ability to observe if the non-malignant BEAS2B cells acquired invasiveness.
Query: Are there strategies to maintain cell viability post PM exposure while ensuring the effects of PM are still observable in the cell line? Would it be advisable to modify the PM exposure conditions, such as by reducing PM concentration and prolonging the exposure duration? Should I provide break for the cell lines to recover and reinitiate the PM exposure?
Hi Jeong Uk Lim
I am working on the effects of seasonal PM 2.5 in BEAS2B cells. Initially my experiments involved determining the cytotoxicity, so i exposed BEAS2B cells with varying doses of PM 2.5 (5, 10, 30, 60, 90, 120, 150 upto 300 ug/ml) for 24hrs. Following exposure I observed that 60ug/ml dose was sufficient to cause a 50% reduction in the cell viability, From the MTT, I selected 5, 30, 60ug/ml dose for the ROS determination, changes in protein expression and inflammatory markers.
* I believe a significant cytotoxicity was observed in your experiment due to 48hrs treatment period and higher dose of PM.
* So If cell viability is to be maintained and invasiveness needs to be studied it would be best to reduce the dose and use a very low dose such as 5 or 10ug, and then a medium dose and further a high dose. This would allow us to study the dose dependent effects of PM and its invasiveness effects.
Thanks,
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