I am working on neuronal cell line SH-SY5Y and glial cell line U-87 MG using 6-OHDA and rotenone as PD stress inducing agents. I am transfecting the cells followed by the treatment. After 24hrs of treatment the cells are dying while remain good in the control group.
Hi,
if you kept the same experimental parameters like before, usually there is a problem with new batch of chemicals or mycoplasma contamination. I would also start a new batch of cells and test it again, because young and old cells could have completely different reaction to the same treatment.
Mateo Glumac Thank you.
Can you suggest any solution for the same?
Mateo Glumac We have done mycoplasma testing and found to be negative for this.
What could be other reasons for such cellular response ?
Shatakshi Shukla
- Changes in chemicals: like if you used all of your old transfection reagent and bought a new lot of the same reagent, changes in chemical that you use for treatment (different lot, polluted chemical, chemical has degraded...). Solution: check if all chemicals are functional and the same like before
- Cell were passaged to many times and are now changed beyond the point that they react differently to the treatment. Also there is a possibility that your cells cot mixed up and you are now using a different cell line all together. Solution: start a new batch of cells and repeat the experiment. I find cell from passage 5 to 18 to be the best for all types of experiments and that reproducibility of experiments between these passages is the best.
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