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My neuroblastoma are floating!! please help


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#1 joycemarievr

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Posted 19 April 2011 - 09:34 AM

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

#2 bob1

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Posted 19 April 2011 - 03:25 PM

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!

#3 joycemarievr

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Posted 19 April 2011 - 03:34 PM

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.

#4 gfischer

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Posted 20 April 2011 - 09:31 AM

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.
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