Hello dear colleagues. I am transfecting 10^6 293LTV cells in 6-well plates with lentiviral plasmids for virus production. I am doing calcium phosphate transfection with 5 mkg of DNA and homemade HBS and Ca solutions. 16 hours after transfection I add sodium butyrate to 2mM. This protocol worked well for about a year giving minimum 10^6 titer of unconcentrated virus. Next day after transfection cells stop growing and change morphology to more rounded cells, which i assume is normal for virus-producting cells.
But recently, cells in some wells become to die in a strange way. Ussualy when HEK293 dies they become round and float up. But my cells seem to lyse while remaining attached to the well, because I see their outlines or shadows (figure B, C). The cells are dying independently of various lentiviral plasmids or calcium phosphate transfection reagents I am using. Ussualy the regions of cell death are varing in size and can affect only a few wells.
Has anyone seen similar HEK293 cell changes? Thanks!
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