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Jurkat cell

jurkat

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#1 carolfrozza

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Posted 09 April 2014 - 10:26 AM

Hi

How can I do to separate live and dead Jurkat cells.? I recently thawed this cell line. I counted with trypan blue and saw many dead cells.

Please, help me !!!

I do not want to lose these cells.



#2 jerryshelly1

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Posted 09 April 2014 - 12:11 PM

Do you have access to a cell sorter? You could also use density gradient centrifugation, but I remember this to be a more harsher method on the cells.



#3 carolfrozza

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Posted 10 April 2014 - 05:47 AM

Hallo, Thank you for your reply. At the moment we have no Flow cytometer in the lab. How does this gradient work, is there a protocol for that?



#4 Tabaluga

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Posted 13 April 2014 - 09:02 AM

Here are some suggestions including a protocol (there are many more on the web, just search for "respective method + protocol" in google)

Insert an "r" into the first link before the "esearch" to make the link work.

http://www.esearchga...cell_suspension

http://www.protocol-...posts/7729.html


Edited by Tabaluga, 13 April 2014 - 09:04 AM.

Il dort. Quoique le sort fût pour lui bien étrange,
Il vivait. Il mourut quand il n'eut plus son ange;
La chose simplement d'elle-même arriva,
Comme la nuit se fait lorsque le jour s'en va.

 


#5 bob1

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Posted 13 April 2014 - 12:30 PM

In addition to Tabaluga's comments, I would like to add that, selecting the cells that are living via these methods is a sort of selection pressure on the surviving cells, which means that the Jurkat cells you now have are no longer Jurkats - so unless these cells are particularly precious (e.g. stable cell line, in which case they aren't Jurkats any more either) then it is best to throw them out and get a fresh batch up.







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