Hello,
I´m currently doing cell culture using Saos-2 osteosarcoma cells . I have only split them 6-7 times, but they seem to have changed their morphology and they do not divide anymore.
I thawed new cells from early stages but the problem continues. I thought it could be a contamination so I discarded the media and everything I used before, but it hasn´t work either.
Any suggestions?
Thanks in advance
Hi Eduardo,
did you check mycoplasma contamination? Even you discard all media and everything, if your cells were contaminated by mycoplasma, you should treat them with antibiotics for mycoplasma. Probably your source of cells were contaminated. Or you use wrong media or conditions for your cell line.
Best
hi,
There are several possible explanations:
(1) The cells are dead because of not being properly stored (problems with liquid N2 container, problems with cryoprotective medium, etc).
(2) Lack of some additives in the medium. E.g. did you add double amount of fetal calf serum? Do your cells need any special additive not commonly used?
(3) For revival, cells were seeded at too low densities. To avoid this, use smaller flasks/plates Why are my cryo-stocks of cell lines not reviving?
Hope this helps
I agree with previous comments and would like to add that SASO-2 cells divide once in 24-36 h and cell proliferation is cell density-dependent. Further, ATCC recommended cell culture medium and culture conditions are supportive of cell growth (see link).
Link: https://www.atcc.org/Products/All/HTB-85.aspx#culturemethod
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