Hello everybody, can someone direct me to a tested protocol to get rid off cell culture contaminants which are resistant to antibiotics (Peinicillin, Streptomycin and Kanamycin). It is importnat that we rescue this cell line as we do not have any other alternatives. Thanks
Gentanamycin will be an option. But becareful: a lot of cell lines, specially fibroblasts, are sensitive to it... My experience has tought me that is the best option for cell lines with hematopoyetic lineage (T-cell, macrophages, etc). What about tetracycline? It could be another option...
Good luck!
We also try gentamycin. Usually 50-100µg/mL. according to literature above 100µg is toxic to mammalian cells. I could get rid of strong contaminants from some epithelial cells and MEFs with genta alone. if cells are happy in genta, then you can always use genta in the medium for maintenance. Another possibility is Ciproflox ( available as CIPROBAY). It is not so well tolerated by cells, but treatment for few days can get rid of most of the contaminants.
You can use the combination of gentamycin and amphotericin B. Gentamycin is 50ug/mL. and Amphotericin B is 2.5ug/mL. It will work 100% confidently.
Thanks Sridhar, but Amphotericin B is antifungal. You mean the combination is more effective in getting rid of resistant bacteria?
I cultured chondrocytes from pig's leg. Penicillin and streptomycin are enough from prevent contamination. Anyway, you have to be careful the tip of pipette and don't forget to pass the fire before close the lid.
Yes Srinivas,
This combination works very effectively we are using this combination for more than 8 years in all kind of culture works including primary cultures and cell lines.
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