I use HUVECs for my experiment, I recently experience problem with culturing them. Apparently it appears it has some bacteria growth in them which appears in their second generation after cultured thaw frozen cells. I've been keen on aseptic technique, changed the media and serum, but still the problem exist. Please can someone advice.
Hi Collins,
Yes, it is definitely possible to freeze down bacteria with your stock cell culture. If this is the case and you've eliminated all other possibilities as to contamination, probably the best thing that you can do is to go back to earlier frozen stocks of earlier passages to see if you have stocks that haven't been contaminated. Otherwise, you may have to start from square one...
I wish you lots of luck,
Rachel
Relative to mammalian cells, bacterial cells are more resistant to damage from freezing as a result of their cell well and ability to accumulate cryopreservative compounds. Many people will culture mammalian cells in the presence of antibiotics, typically penicillin/streptomycin, to kill contaminating bacteria in cultures. The antibiotics will not harm mammalian cells.
I had the same experiments with yeast contaminations. certainly, yeast, bacteria and mycoplasma survive very well after freezing in liquid nitrogen. Wash your cells by repeated antibiotics treatments.
You can definitely freeze down bacteria - look at the way stocks are kept for molecular biology work!
If you don't have any earlier stocks, split your cells into around 10 different TC dishes and try other antibiotics at a range of concentrations - your bacteria must be resistant to the antibiotic you routinely use in cell culture. I assume you are using an antibiotic to limit infections.
- Badges
- Science topic