Cell culture tips and tricks

• Bring the temperature of the cell culture media to 37^oC in a clean water bath, preferably dedicated to cell culture use only, to decrease chances of contamination.

• Keep the biosafety cabinet (the cell culture hood) empty, except when it is in use and stay clear of the vents to promote proper air circulation. This decreases the chances of contamination. Wipe down the work surface with 70% ethanol before and after using the biosafety cabinet to maintain a sterile/ an aseptic environment.

• Spray and wipe down everything (pipets, media bottles, other conical tubes, tube racks, etc) that enters the biosafety cabinet with 70% ethanol, to reduce chances of contamination.

• Refrain from placing caps, pipets, or other equipment down on the sterilized surfaces. If a lid needs to be opened, be swift in keeping bottles and flasks open, and quickly close them to reduce the time the bottle stays open to potential contaminants. If caps and lids must be placed onto the work surface, briefly place them open-side up, and ensure that nothing touches the inside of the cap/lid.

• If you suspect a pipet tip may have touched a surface (even if it has been sterilised), dispose of the pipet tip immediately and start with a fresh tip. The contact, albeit brief, could introduce contaminants (e.g. bacteria, fungi) that could exist on surfaces (the biosafety cabinet walls, pipets, cell culture dishes/flasks) into the cell culture medium, the cell stock itself, or any other supplements and addition stocks.

• Unopened cell culture media can be stored in the cold room or a 4 ^oC fridge, and should be used close to, or before the expiration date. Once supplemented with FBS and/or other additives, the media should be used within 6 weeks of opening.  The supplemented media can be stored at 4^oC during this time, and warmed to 37 ^oC before using.

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