Contamination of cultured cells - (Nov/01/2007 )
So today I went back to lab and found out my cell line was contaminated again. And I dont really know how to handle this situation. Coz this is already the second time in a roll ofcontamination, it happened as well last week. What do you guys usually do once your cell line was contaminated?Also, how to minimize the chance of contamination?
Thanks
Dear Travis,
About incubators
RS/Binder/Nuaire/Heracell have high temperature decontamination cycles which aid cleaning of the incubator. These are different temperature cycles: 120/180/90and 140/90 oC respectively.
This does not however solve the problem. As many have said USE GOOD ASEPTIC technique.
Have a rota for all things tissue culture:
Weekly changes of water in waterbaths/Incubators
Stocking of TC rooms
Cleaning of Floors and work surfaces in TC areas.
Have seperate side labs for TC work
Have seperate labs for different applications i.e. Viral transfection, primary, human samples, Mycoplasma negative and quarantine areas.
Cleaning of class II cabinets.
Making up IMS
Have seperate TC lab coats
Use quality TC ingredients
USE PLENTY OF IMS to wash gloved hands. Use IMS to clean TC flasks, in and out of the CO2 incubator.
GET TRAINED BY SOMEBODY WITH PLENTY OF EXPERIENCE.
CHUCK AWAY ALL CONTAMINATED CELLS......DO NOT TRY TO CLEAN THEM UP.
DO REGULAR MYCOPLASMA TESTING.
Some people are "luckier" than others with contamination..........these people always have better technique than those who are "unlucky".
And as Dominic rightly said.....these observations are from 30 years of experience.
OH!
Another issue I see often is technicians storing the pipets, tips, buffers, biohazard bags, and other commonly used items inside the BSC. It is never a good idea to store any items in the hood as this not only prevents effective cleaning but it also blocks the air flow inside the hood.
We have several Heracell incubators in our facility and the "sterilization cycle" is not intended to replace a monthly manual cleaning. If the cycle is run too often the seals on the sensors will go bad. Which is a serious issue with the water sensor at the bottom of the incubator because the water will leak out. But it is very helpful in preventing the spread of contamination.
[quote name='Tsunade Sai' date='Nov 7 2007, 06:02 AM' post='116187']
thanks for your advices! ter are really helpfull!!