Hello
I have these human fibroblast cells in culture for a while and they look fine with a typical fibroblasts morphology, At passage 8 the culture looks like a heterogenous culture with different cell morphology (pictures in the attachment). I am not sure what the reason is or if it becasue of cross-contamination with other cell line which I think is unlikely because I always work with one type of cells at a time.
Hi.
I have noticed the same situation. I work with MRC-5 cells which are lung fibroblasts. At early passages, they maintain their normal morphology. As you continue to passage them, they grow old and for some reason (My guess all the constant treatment with trypsin and antibiotic), they become vacuolated and acquire spheric morphology (I noticed this around the 13-15 passage) and indeed, it affects the result of the following treatments I have to perform. So I decide to thaw new cells and start with optimal cells.
It might be mycoplasma. You can get this in your tc from doing rodent work same day you use the hood or from cheap fetal bovine serum. You can test for this intracellular menace using pcr
Worked with fibroblasts for a long time. Could be several things. From infection, use of different batches of trypsin to old age (high passage number) and so on. Once you observe that, its best to change to a lower passage number as well as test for infection. Otherwise fibroblasts are one of the easiest cells to culture. Good luck!
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