Cell fusion protocol to generate heterokaryons - (Jan/26/2005 )
Hello !
I'm trying to generate heterokaryons from 2 cell lines, namely U937 (monocytes) and Hela cells. I'v tried different protocols and used commercial PEG from Sigma but I got only 3% of fused cells and which don't survive after FACS sorting. Problem is : one cell line is adherent and the other is not. Has anybody had the same problem and who could help me out on this ?
Thanks !
Alex
-osiris-
QUOTE (osiris @ Jan 26 2005, 11:08 PM)
Hello !
I'm trying to generate heterokaryons from 2 cell lines, namely U937 (monocytes) and Hela cells. I'v tried different protocols and used commercial PEG from Sigma but I got only 3% of fused cells and which don't survive after FACS sorting. Problem is : one cell line is adherent and the other is not. Has anybody had the same problem and who could help me out on this ?
Thanks !
Alex
I'm trying to generate heterokaryons from 2 cell lines, namely U937 (monocytes) and Hela cells. I'v tried different protocols and used commercial PEG from Sigma but I got only 3% of fused cells and which don't survive after FACS sorting. Problem is : one cell line is adherent and the other is not. Has anybody had the same problem and who could help me out on this ?
Thanks !
Alex
How does PEG help in fusion protocol.
-MUTHURAMAN-
PEG causes dehydration of membranes and promotes membrane fusion.
-genehunter-1-
I know inactivated Sendai virus are much more efficient in mediating cell fusion.
-genehunter-1-
hi,
i used to use, 50% PEG 4000 and 10%DMSO for fusion of cells, wat i learnt was, it is not advisible to use higher polymers like PEG 10K or more.
gud luk
payeli
-payeli-