working with THP-1 cell line, and even my culture media gets contamination. I'm trying of doing quality control to everything, but I don't know what else to do..
Test your media and all of its components (penstrep, FCS, etc) by putting a small amount of each in individual wells of a 6 well plate and incubating for several days. Look for contamination daily to identify which component(s) is contaminated. Once you've identified this, get a new sterile bottle of that component(s). Sterilize your incubator by autoclaving the shelves and cleaning the inside with ethanol or Lysol I.C. Quanternary Disinfectant before starting fresh with newly thawed cells in newly prepared media.
Please see the following links:"
https://ocw.mit.edu/courses/biological-engineering/20-109-laboratory-fundamentals-in-biological-engineering-spring-2010/labs/guidelines-for-working-in-the-tissue-culture-facility/
https://www.youtube.com/watch?v=PQ1ESPYnI2w
Do you know what kind of contamination your are getting? Many bacteria will look like squiggly oscillating rice under the microscope and yeast will be apparent by a smell in your incubator.
Overall, make sure to have good hood etiquette to make sure that you are protecting your cultures from yourself. Make sure to maintain the recommended sash height and depth into the hood itself, make sure to not pass your hands/tools over open media containers or cell tubes, things like that. Then, in addition to your incubator maintenance and your medias, if you are still having problems I would recommend carefully disassembling your culture pipettes and cleaning them with ethanol, wiping down and cleaning the hood components and under sash with ethanol, replacing filters in serological pipettes and vacuum catch flasks, making sure to clean vac lines regularly (I run 50% bleach through my line before and after use followed by a little DI water to rinse the line), and making sure any eppendorfs, tips, etc are sterile/autoclaved appropriately.
These helped me a lot, and youtube and this https://www.thermofisher.com/se/en/home/references/gibco-cell-culture-basics.html
All manufacturers provide free and good information on how to perform different techniques. They are interested in you doing it correctly and working with their products as intended so you don't later blame them and buys from their competitors instead!
Kindly give a sterility check to your complete media for overnight at 37oC before using for culturing cells.
Check what kind of serum you are using. Do you heat inactivated it or filter sterilization. Try to work with aliquoted, so that chances of contamination are minimum
Thanks! Evereybody! Boaz, for the links; Kressling for the tip of the waterbath, Henrry I really need to take care my cultures from myself!! Rasmus, I saw the videos before, thanks for the links. And Harsh I started to chek in complete media because I was anly doing a check with agar LB :C
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