Hello, I am working on human dermal fibroblasts. After thawing from liquid nitrogen, there is clear morphology till one/two passages. But after that we observe a change in morphology, like the cells have been damaged. Please suggest the reason behind this type of behavior. I am attaching the images of the healthy Fibroblast and the damaged one.
Hey Bikash,
Looks to me like a intra-cellular attack of a microbe. Notice them within the cytoplasm, they appear granular.
Best,
AMB
Hi Bikash
I agree with Abdul Mannan Baig, you culture is clearly contaminated.
you should discrad all your stock and get some new ones, and try to track you source of contamination.
Best wishes
Daniela Fabrino
I agree with my colleagues. You seem to have an infection in your cultures.
Change everything you used (medium, dishes,...) and restart a new culture. Good luck
Hi Bikas,
I agree with the others. your cells have a heavy load of contamination.
check this paper for similar problem that you are facing https://www.ncbi.nlm.nih.gov/pubmed/19926304
discard all the vials, medium, PBS and evrything. I have observed that this starts because of contaminated water being the source. Autoclave and double sterilise the reagents you use for culture.
I agree with them about the possible contamination being the cause for your cells to look like they do.
These are my suggestions:
(1) Try to thoroughly clean the surface of skin before enzymatic digestion of the specimens.
(2) Determine the source of contaminations, such as mycoplasma, at each passage of the primary culture.
COLLEAGUES ARE COMPLETELY RIGHT ONLY infection can lead to change the appearance of fibroblasts,. and this is from my experience and work before! And best once again to the dishes before use to disinfect and clean, so as not to be repeated!
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