I am using colon cancer cell (COLO-205), which is a mix-type of cell (suspension and attached), if I take the media and centrifuge it and then I resuspend the cell with media, and then I add it again with a new media. Is this subculture or a change of medium?
And how can I change the medium for complete cell suspension?
Dear Bashir,
If you centrifuge the cells and after removing supernatants, re-suspend in fresh medium, it would be changing of medium; but if you divide it in new flasks it is the subculture.
Hello Bash,
Centrifuging and re-suspension of your cell with an entirely new medium is changing of medium. Subculture is introducing some of your original culture into a fresh medium.
hello basheer,
here what you are doing is splitting of cells. that is you are dividing your total number of cells into two flasks. this is what we call splitting of cells. for changing the you need to wash the flask with 2 ml PBS(phosphate buffer saline)
Protocol: discard the medium
add 2 ml of PBS and flush it with the pippette and then discard the PBS
then you can add 5 ml medium.
this is the way to change the medium, if you dont have the composition of PBS you can ask me i will give you.
with regards,
Muneeb Hamza K.H
1: Collect liquid and spin down to get pellet
2: discard supernatant, add PBS, re-suspend, and then spin down to get pellet
3: during that time fill next flask with appropriate amount of media
4: discard supernatant, re-suspend with appropriate growth medium and transfer appropriate volume to new flask (already filled with fresh media in step 3)
Hi
I have a related question to the above question.
For suspension cells, if you have multiple cell plates and change the media for all the cell plates. Suppose you combine cell solutions from all plates into one centrifuge tube and re-suspend in the fresh media. At this time, will you change the Passage Number, or will it remain the same?
- Badges
- Science topic