There is no contamination in cells. passage number is :30. Still after every passage they look more elongated. I use RPMI 1640 medium with 10% FBS.
Most epithelial cell lines under certain stress change shape. Therefore, thawing another vial may not help you. You may want to identify the source of stress under your culture conditions. To start, have an another cell line that under similar culture conditions does not change the shape (control). See if your medium is low in glutamine or other essential growth components. Serum source can be a contributing factor.
Hello Hina,
Why not thaw out some low passaged cells and use for your studies?
Most epithelial cell lines under certain stress change shape. Therefore, thawing another vial may not help you. You may want to identify the source of stress under your culture conditions. To start, have an another cell line that under similar culture conditions does not change the shape (control). See if your medium is low in glutamine or other essential growth components. Serum source can be a contributing factor.
_the things discussed about trypsinization time, cell density requirement for maintenance or lifting cells too early once trypsinize (within day or 2), change in culture vessel?? after taking all care about these aspects; still have you facing the same problem?
i agree with Divaker. You may also try to thaw lower passage (p0-p4) of A549 under same condition and compared with the current A549 with p30.
It is advised not to use higher passage for your experiments. Try to maintain lower passage as conducting your experiment.
Thank you Nazilah mam. I will keep in mind all the important points. Thank you.
I have had the same characteristic. It is not very clear which factor has actually the impact to transform the cells morphology. It's well-known, however, several passages will have detrimental genomic changes, that might includes decrease/increase in the chromosomes number.
consider contamination of your culture with different cell type...
Hello Hina check your medium. RPMI probably is not the appropriate medium. Try the medium recommended by ATCC for this cell line. Hopefully, Your problems may sort out just by doing that. Please post back if it improved by doing that. Best wishes
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