how to seperate cells from the dead and the live - (Oct/29/2006 )

I have k562 cells on my hands. Severals days ago I forget to reflesh the cell culture until the liquid has turned yellow seriously.

Actually I personally think it is possible to recover the cells if you only transfer minority of them.
But my plan fails this time. Most of them are still shrinked.

Of course I can get new resurgent cells. However, It makes me think about a very interested question: If it is possible to seperate the dead cells and the lives by some simple methods like one-step centrifugation? (just obtaining the most the lives is my purpose.) and which part will they locate: the bottom or the top?

Hi Rosalind,
you not here tell anything about nature of your cells like adherent or suspended growing cells. In the case of adherent culture dead cells flowting you can removes its by removing media and addeing fresh one. In the case of suspension culture spin culture at very low speed at which only live cells able to pellets , here you not able to getting 100% live cells but you can increse its % in the cells population

We use MACS Dead cell removal kit. I dont know how that works. If anyone knows please share.

thank you for your advice! these are very helpful!

Hi everyone, I've just started work on K562 cells. Any idea how we can detect contamination in a Suspension cell line?

Can anyone help? So far, my solution is orange in colour (after changing the media) and i observe small black spots on the bottom which i've been told are dead cells...

Here's a link.

http://www.miltenyibiotec.com/en/PG_712_10...emoval_Kit.aspx

You can use Ficoll to separate dead cells and live cells. Its very simple. you can refer to the book 'culture of animal cells' by Ian Freshney.