4 replies to this topic

[carolfrozza]

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.

[jerryshelly1]

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.

[carolfrozza]

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?

[Tabaluga]

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.

[bob1]

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.