U87 cells
Jennifer Robertson
however the ATCC site doesn't give much information about them. Mine seem to
clump before even reaching 50% confluence. Even when I suspended and
replated them at about 70% confluence the cells settled and clumped again.
Is this normal for this cell line? I know i'm seperating the clumps well as
I triturate the begeezus out of them and I can see that its all seperated.
Any input would be appreciated.
J
Emir Khatipov
not mistaken, they are derived from brain cancer), and clumping of cancer
cells is quite common. LNCaP cells (prostate cancer) that I work with do it
al the time. There are few things I could recommend you to try to see if the
cells start clumping less. One thing is the volume of the medium you grow
the cells in. Try using more media. More media means longer time before the
cells acidify it. High acidity is one of the unfavorable growth factors, and
it will cause the cells stick together to survive. You might check if your
cells will grow on other higher buffered media, or check bicarbonate
concentration in the medium you use and CO2 concentration in your incubator.
Third thing, your cells might be less prone to clumping in richer media
(e.g. RPMI vs. DMEM). Another thing, you may try using plasticware coated
with poly-lysine, collagen, etc. for increased binding of the cells. The
stronger the binding of the cells to plastic, the flatter cells would
spread.
And don't treat them too hard. You might even be able to flush them from the
plastic without tripsinization. In my impression, even if you don't brake
the clumps before plating, the cells will spread on the plastic anyway (and
then start forming clumps).
- Emir
"Jennifer Robertson" <jrobe...@sympatico.ca> wrote in message
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Araluen Freeman
Jennifer,
I've cultured myelocytic leukaemic HL-60 cells. They're a suspension when
happy, but the one time I passaged them into IMDM that hadn't been
supplemented with FCS and antibiotics, they stressed and clumped. Perhaps
the media isn't the flavour of choice for U87?
G'luck.
"Jennifer Robertson" <jrobe...@sympatico.ca> wrote in message
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Leman
>news:9cFs7.9945$%r.28...@news20.bellglobal.com...
>> Has anyone ever cultured U87 cells? I think mine are behaving strangely,
>> however the ATCC site doesn't give much information about them. Mine seem
>to
>> clump before even reaching 50% confluence. Even when I suspended and
>> replated them at about 70% confluence the cells settled and clumped again.
>> Is this normal for this cell line? I know i'm seperating the clumps well
>as
>> I triturate the begeezus out of them and I can see that its all seperated.
>>
>> Any input would be appreciated.
Jennifer,
U87 just happened to be my "favorite" line, and I had the same concern
when I started working with it two years ago. This behavior is so
different from other GBM cells, that I started worrying and asking
people from different institutions to send me a vial of their "breed"
of U87.
ALL of them start growing spheroids once they are a little more than
50% confluent. The only thing you can do is not to plate more than
5-6M per 100-mm dish and use them for your experiments within next
couple of days. They are also very hard to stably transfect, since the
working concentration range of selection agents is very narrow. For
example, you might not get much selection at 200 mkg/ml G418, but
completely wipe out the whole dish at 500 mkg/ml. There is a very
narrow window in between, in which you'll get nice colonies.
Anyway, to make a long story short: yes, U87 are just strange and
don't quite behave the same way as other GBM lines.
Good luck with your studies,
Leman