Hi all,
I made a mistake for the first time during my 7 years of cell culture and I have no idea what I should do. So I just made a fresh cell culture media stock in the original bottle (Gibco DMEM + 10% FBS +1% Pen-strap). Then I use this stock to subculture cells and plate them in petri dishes as usually. However, I wasn't focus enough and I accidently use a pipette tip that has contacted diluted cells to transfer fresh media from the stock bottle. You can imagine there are cells attached to the wall inside the pipette tip and now they are floating in my 500ml fresh stock. I took out 3ml from the contaminated media and plate in a petri dish to test if they will grow. I will be using the media for the same cell line all the time. Should I throw it away? Or I can still use it on the same cell line? Since its worth over 60 dollars (cause it has 50ml FBS in it), I hope I can still use it.
Any idea is appreciated!
You could filter your medium through a 0.2 micron filter (which will remove both bacteria & human/animal cells). Or if your contaminated pipette was still sterile when it went back into the media bottle you could centrifuge the DMEMit to pellet the cells, not as clean as filtering but cheaper!
Hi,
If the media contains phenol red it will turn pink (basic) or yellow (acidic) depending on the change in pH. So until it remains red you need not worry about it. Since you have added Penstrep chances of bacterial contamination are less. However you can conduct a sterility check by inoculating it on Nutrient Agar plate.
Thank you.
You could filter your medium through a 0.2 micron filter (which will remove both bacteria & human/animal cells). Or if your contaminated pipette was still sterile when it went back into the media bottle you could centrifuge the DMEMit to pellet the cells, not as clean as filtering but cheaper!
Hi,
I agree with Mark and Jaison basicly. Also you can filtered the whole medium , filtered via 0.2 micron sterile filter is the best way. But the cheapest way is incubating the Medium at -20^C (freezer) for only 2 hours. The cells will be die but, when you use it, you can centrifuge the medium 600 x g (high speed) for 5 minutes.
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