My cells namely Kasumi-1 and Thp-1 previously was healthy and growing at a steady rate but just a few days ago it appeared to have been contaminated with the same bacteria that plagued me previously, Acinetobacter. I have checked if the media is fine and I sterilize all pipettes, flasks, tubes and etc before ever started my work to prevent any contamination. Everything is sprayed down with 75% ethanol before I begin culturing my cells so the only other explaination is the incubator that is contaminated but I have 2 other colleagues who have their cells in the same incubator and is not contaminated. With the cause of contamination aside, how can I get rid of Acinetobacter and save my suspension cell lines? I have no kits to remove contaminants, I only have Pen/Strep. I have attached an image of Kasumi-1.
Hello Aswani Jaishanker
You can try using 0.2% Normocin from invivogen. It can be added to the culture medium in combination with Pen/strep to broaden the anti-bacterial spectrum.
Good Luck.
Hi Aswani Jaishanker
Split your cells into 12-well/6-well plates and add your antibiotics at different concentrations (usually 10-fold). Determine the maximum concentration that your cells can survive in and then culture in these conditios with the antibody for a few passages. After this, your cells should be contamination-free (althought check this first!) at which point you can freeze stocks.
Hope this helps!
Jack Hopkins Thank you so much, this really helps. I will definitely try this.
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