3 replies to this topic

[joycemarievr]

Hi all,

I have cultured the SK-N-SH and the SH-SY5Y cell lines for many years in grad. school.  I never had any trouble when thawing them, but now in a new lab they stay floating.  I have centrifuged it after being 24 hours in the incubator but it has not attached.  I am doing all the same i used to do before, but can't figure out why cells are not attaching to the flask.  I already tried changing the size and bran of the flask but the same happens.  What other posibilities might be causing this?

Thank you,
joyce

[bob1]

Are the cells dead?  A quick look at them with trypan blue will tell you.

Sh-SY5Y have a mixed population of cells with some being adherent and some floating, both should be cultured!

[joycemarievr]

thanks.  yes they are alive.  but not even one cell has attached.  I used different kinds of flasks and check the CO2 levels but nothing.

[gfischer]

If people in your new lab had previously not cultured the cells correctly, and only cultured the non-adherent cells, they may have selected for that population.  If this is the case, you should probably get a new batch of cells.