I have noticed these cells which seem considerably larger than the majority of the cells in my tissue culture flasks for NCTC clone 929 and LN-229 cells.
The media for the flasks looks normal and isn't cloudy. I initially thought it was some sort of contamination so I prepared new media, threw out the flasks and started growing a new batch of cells which I had previously sub-cultured - it's possible that these are contaminated too but they were frozen a considerable time ago when I hadn't noticed this strange cell morphology.
I have attached some pictured of my NCTC flasks. In two of the pictures the cells are detached and the difference in cell size is very evident.
Can anyone suggest what these large cells are?
Hi
In your images
1. cells with distinct membrane are may in a attaching or detaching condition. Mostly after trypsin treatment of over confluent cells can show this morphology
2. Another large cell with distorted membrane are going to burst due to change in osmosis
Hi,
May be due to your frozen technique. The cells are not looking healhy due to some toxicity. please check the following papameters
Thanks for the answers
Mamatha - I agree that the distinct membranes are due to the cells being in the process of attaching or detaching. Are you saying that you think the difference in cell size is due to overconfluence?
Deepa - The cells are stored at -80 °C and the temperature of the incubator is correct. The cryomedium used was 10 % DMSO 90% serum and on revival 10 % serum is used in the media. The majority of the cells in each flask look healthy and I have to spend a while looking for these large cells as they are few. I am mainly worried that the large cells are due to cross contamination with another cell line - do you think that they are larger due to toxicity possibly from the cryo conditions?
Thanks
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