QGP-1 cell line  maintenance protocol 

Warm media

Swirl cells in 37 degree water bath until vial is thawed

Add 7 mL of culture media to a T25 flask and gently pipette thawed cells into flask

Let cells adhere for 24 hours and replace half of the culture media with fresh media

Replace the media in the flask with fresh media after 48 hours

Split cells once cells reach 70-80% confluency: usually takes 3-4 days from thawing and plating in a T25 until they are ready for passage. (they won’t ever become completely confluent, they will just start to grow on top of each other)

Lift cells using trypsin and plate all cells in a T75

They are usually ready to passage 3 days later.

Warm media and trypsin in water bath

Place flask back in incubator and split next week

Clean hood with EtOH

Remove media

Wash with HBSS

Discard HBSS, add Trypsin, and place flasks in incubator for 5 minutes

Bring flasks back to hood, obtain lifted cells and spin at 500 rcf for 5 minutes

Discard supernatant

Resuspend in 10 mL

Place 250 uL of cells into a T75 flask and add 13 ml of media