non-viral transfection of MSCs
Posted 27 September 2004 - 02:38 AM
I have read few papers that successfully transfect MSC with Lipofectamine, calcium phosphate, Amaxa Nucleofector and Eppondorf Multiporator. But I failed to see any glowing cells (my construct is pEF-X-IRES-hrGFP) by using these method (expect Eppondorf Multiporator, I haven't tried it yet). Should I tried more different conditions with these have-been-succeeded methods? Or non-viral transfection of MSC is really so tricky thus I should switch to retrovirus systems or so? (Btw, what are the advantages and disadvantages of adenovirus, AAV, and retrovirus? I do not know how to choose...)
Is there any one having experience with it and can give me some advice? I would very appreciate it !!!
Posted 27 September 2004 - 09:34 PM
could it be that your vector is not activated when transfected? Are you sure that it works, did you see expression and glowing with it somewhere else?
Posted 30 September 2004 - 05:24 AM
Is that what you meaned?
Thanks anyway !!!
Posted 30 September 2004 - 08:52 PM
yes that is what I meant. OK, so have you tried to transfect these cells before, with someother vector? Just to make sure that it's a problem with transfection...
Posted 02 October 2004 - 08:49 PM
Posted 19 October 2004 - 07:09 AM
so i think it's not because lipofectamine2000, but your vector or cell, be sure they are work well.
Posted 15 November 2004 - 03:27 AM
I have very good experience with the amaxa nucleofector and the gene transfer into hMSC, 40-70% are absolutely no problem. I think the main problem is the vector, although your construct does work in 293 cells, it could be that the expression pattern in primary cells is different.
A friend of mine, Dan Gazit from Israel, is promoting this technology for non-viral tranfection into human MSC on his homepage. He is an opinion leader in tissue engineering, perhaps you can find more clues on his hompage:
Just search with google : Dan Gazit, mesenchymal stem cells