I am using HeLa cell for my studies and i am retriving the cells but it wont attach. the unattached cells are creating the extra bubble like structure and it contains the movement particle in that. You respected people have any words or ideas for me... i m waiting for valuable suggestions.
Hello, Devan.
Sorry to tell Your cells may be contaminated judging by what you wrote and Fig uploaded. The empty vesicle-like structures are lysed cells swollen resulting from cell freezing or thawing procedure. The moving paticles appear to be bacteria or some other microbes. Please use a newly bought cell stock.
Best wishes.
Hello, Devan.
Sorry to tell Your cells may be contaminated judging by what you wrote and Fig uploaded. The empty vesicle-like structures are lysed cells swollen resulting from cell freezing or thawing procedure. The moving paticles appear to be bacteria or some other microbes. Please use a newly bought cell stock.
Best wishes.
These balloonizations that you describe are usually observed on lymphocytes in culture, and are due to an infection by mycoplasmas. These bubble-like structures anticipate cell death. It was thirty years ago these ballonisations were regularly observed during infection of cells by HIV, and represented a test for infection, but this was due to the HIV-infection by strains of mycoplasma. I suggest you change cellular source, and check where does the contamination before continuing with HeLa cells.
Thank to you all for supporting me...and i have another doubt.
can contamination come through DMSO while we r storing the cells?
Hi, Devan.
It is unlikely. DMSO is a potent toxic solvent being used as a cryoprotectant with serum. Probably, contamination may come from something else, including serum, medium and non-sterile procedures.
Regards
Sang Ho Lee May I ask you to back up your statement with a scientific paper so that I can read more about it? Thanks
Hello Mehri,
I am sorry, but no such a paper. Because I mentioned many possible things that we all often come across, careful reading of any books or reviews on 'animal cell culture techniques' and 'cell death' would help you avoid such mines above. There are too many must-do and do-not in preparation of medium, making aliquots of stocks for convenient use, followed by storing at -70C until use, freezing and thawing cells, washing with appropriate volume of fluid to eliminate any possible particles or microbes at a proper speed.
And then, good practice and sharp eyes are also required in cell culture lab with various types of cells. Long hours of cell observations during cell thawing, washing, plating, dissociation, expansion, freezing and etc., too.
Although I do not think you are a novice, here' are links below;
Cell culture tech
https://www.vanderbilt.edu/viibre/CellCultureBasicsEU.pdf
https://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma-Aldrich/General_Information/1/fundamental-techniques-in-cell-culture.pdf
https://www.cpp.edu/~rmogul/safety_files/Aseptic%20Technique.pdf
https://pdfs.semanticscholar.org/a9f1/0fc74845a1491676c61acc1887557d684d70.pdf
Cell death
Article Morphological Features of Cell Death
https://www.intechopen.com/books/cell-death-autophagy-apoptosis-and-necrosis/necrosis-as-programmed-cell-death
https://www.nature.com/articles/s41392-018-0018-5.pdf
Best regards.
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