Subculture of Adherent Cells

[@joseluisbioq]

PBS is a buffer with a high salt content, if you put it on 37C for too long it will precipitate and change its concentration. Trypsin is removed in orden to minimize the required amount of serum needed for neutralisation. It also helps to reduce the stress given to cell when they are detaching

[@Delphinolinguist]

Be very careful if you decide to aspirate the trypsin off. You need to be familiar with the particulars of the culture of the cell line you are working with. With cell lines that detach very easily, you could end up aspirating off most of your cell mass if you decide to aspirate the trypsin even after half a minute of contact with the cells. With some lines this will not be a problem, but it's important to know whether it will be a problem for your cells.

[@bharanitharan3997]

instead of aspirate trypsin and remove most of the cell mass along with it, you can centrifuge cells along with trypsin, aspirate the trypsin, add media to centrifuged cell pellets and then add it equally to 2 flasks.

[@wayne02058]

i rewatched the video and i assume that the trypsin was only in there and had not enough time to dislodge the cells from the flask, so if she works quickly she should be ok since she has 80% confluency. and your method does seem to have the potential to save cells from being aspirated away, especially when working with an expensive cell line.

[@larissamarina1291]

We do the same here

[@funny11744]

BUt if the trypsin did not have time to act - your proposal ....not good. We can lose many cell.

[@mercurypark]

I wonder why aspirate off most of trypsin. Is it for reducing cell amount? or removing some cell that condition is not good? Answer me please...

[@JNNT934]

The thin layer of trypsin is enough to detach cells from flask or plate.

[@sophannarykong2320]

when you removed most of the trypsin, how long did you put the flask in the incubation room?

[@PhilWithCoffee]

It depends on the type of cells you're growing, but it's standard to let it sit for 1-20 minutes. You just have to check it every so often to see how well the cells are breaking up (and gently hit the sides to encourage it).

[@misbashaikh7646]

2 mins.. depends on cell type

[@12823matthewkao]

wait....so you no centrifugation is involved? wat.... this demo is kinda wrong in many ways

[@mohsentaghavi8063]

Why you don't put D-PBS in 37 C water bath ?

[@OfficialNetDevil]

Can this be done to stem cells?

[@muskansheba6779]

wht is the passaging ratio done in this video?

[@tingdarren840]

i also want to ask. Do you get it?

[@lingcheeshuen9183]

I also want to know it

[@cosmicconstellations511]

I think it is 1:3

[@keyvansobhani4644]

No centrifuge?

[@rillrill1312]

this beat is fire

[@asmaalwerfaly4776]

Thank you this is helpful 🌸

[@misbashaikh7646]

Wjy do we add pbs

[@kosheeka]

PBS enables us to do the following in our cell cultures: • Prevent osmotic stress on cells • Maintain physiological pH • Facilitate washing and dilution procedures • Provide a basic transport and storage medium

[@mygan1010]

the music is freaking creepy.

[@mrbayardi1]

makes subculturing interesting to watch lol