Jun 23, 2022
QGP-1 cell line maintenance protocol
- 1mayo clinic
Protocol Citation: bellampalli.shreya 2022. QGP-1 cell line maintenance protocol . protocols.io https://dx.doi.org/10.17504/protocols.io.bp2l61e1rvqe/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: June 23, 2022
Last Modified: June 23, 2022
Protocol Integer ID: 65196
Abstract
QGP-1 cell line maintenance protocol; thawing and passaging QGP-1 cells
Thawing protocol
Thawing 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.
passaging protocol
passaging protocol
Warm media and trypsin in water bath
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
Place flask back in incubator and split next week