I am facing problem with cell culture, after I start the culture, the media turned turbid in about 2 days. I tried everything but the problem still persists.
Under the microscope the cells look ok and no change in morphology.
Does anyone have an idea about this ?
I am working with Hucct1 and HepG2 cell line.
Hi, It seems to me that your cell culture is contaminated. Do you use antibiotics in your medium?
It seems that your cell line/your media is contaminated. Prepare all the solutions viz. Trypsin, media, PBS freshly and decontaminate your cell culture lab with proper method and thaw a new uncontaminated stock of your cells. I assure that you will not face this problem again.
your cells may be contaminated even if they look ok under the microscope. The only thing you can do to ensure decontamination is to change all the stock solution and thaw or buy new cells. You may also decontaminate the cell incubator.
Hi Brunette, as Nathalie said I will decontaminate first your incubator. In particular if you have incubator with Hepa filter, and you have contamination with bacteria even if you thaw another cell stock you may caused contamination of the new one (personal experience). It is better that you decontaminate the incubator, change all your media, serum, trypsin or any other solutions that can be contaminated, use anly filter tips and then thawn your new cell stock.
The culture is probably contaminated, get rid of everything and review your procedures especially if you do your own culture media (by filtering etc, I prefer to buy ready to use)...decontaminate incubator and use sanitizer instead of alcohol (more effective and cheap) I suggest LPH or Coverage Plus
It is probably a contamination. You should test everything you use with this cells for contamination (media, PBS, FBS). First you should find the source of the contamination and then you will know how to deal with it.
Sounds like a contamination problem. Even though the cells may appear 'normal' they may actually not be. Others have very rightly pointed out that you need to decontaminate the incubator and all the equipment (pipettes, etc) that you use and I would get a fresh batch of cells, PBS, media and trypsin/accutase and any other reagent that you use. It is possible that one of these may have been contaminated. All the best.
Dear,
The turbidity might be due to the contamination. If you are no using antibiotics in cell culture media, it is better to supplement the media with antibacterial and antifungal antibiotics in optimum concentration that will not affect the health of the cells.
in general, Penicillin and streptomycin are used as antibacterial antibiotics. Amphotericin-B is used as antifungal antibiotic.
Antibiotic-Antimycotic solution (100X) is available from Sigma-Aldrich. This can be used directly at a required dilution given in specifications.
Apart from this, ensure the cell culture cabin (laminar chamber), apparatus... to be aseptic.
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