Problems when plating neuroblastoma cells
Posted 30 March 2010 - 04:09 AM
I am trying to do IF with neuroblastoma cells (SK-N-SH) using our "traditional" protocol, which works fine with HEK, COS and so on.
We plate cells on 12-well plates with a sterile, poly-lisinated coverslip inside, and then perform transfection or IF.
Usually when we plate the cells on such coverslips, cells grow well and only the cells affected by the movement of the coverslip around the well die.
My neuroblastoma cells look healthy when plated on larger flasks with good confluence, but when I tripsinize and plate them for IF, occurs a massive death and surviving cells look terrible. If I use non-lisinated coverslips, it is worst. Transfection efficiency with lipofectamine is poor.
What can I do? I try not to plate them at too much low confluence because they are very sensible.
Posted 30 March 2010 - 08:32 AM
Posted 30 March 2010 - 11:28 PM
Yes, the coverslips are coated with poly-L-lysine...
Posted 05 April 2010 - 10:35 AM