Roller-bottle cell-culture systems are widely used for the production of proteins or viruses.
To generated the production in the cells transfection of one ore several plasmids is
needed to express the appropriated gene to generated the desired product.
Transfection in large cultures vessels and roller bottles is expensive in labor and media
needed for the culturing of the cells. Finally the general outcome of the transfection
experiments shows only 10-15% of the input cells are transfected and massive cell-death is
induced due to the transfection protocol.
Therefore Synvolux developed a product application of SAINT-MIX for the efficient transfection
of roller-bottle production cell-lines, like CHO-K1, HELA S3 and HEK293.
Transfection efficacies up to 70% and cell death lower as 10% can be achieved.
Expression has been visualized by Guava –easycyte (Facs) analysis and is detected already
at 24 hours after transfection.
Guava-easycyte analysis of Hela S3 cells transfected with SAINT-MIX.
GFP expression is measured after 72hr.
Repeated transfection of plasmids encoding for different genes, as needed for viral production, have been showed by sequential transfection of plasmids encoding GFP and dsRED.
Transfection of 1 ug GFP (green bars) and 1 ug dsRED (red bars) are set to 100%.
After transfction of dsRED plasmid the GFP plasmid is transfected after 1 hr of recovery of the cells. The upper 3 bars groups show 3 different amounts of dsRED combined with 1 ug GFP plasmid.
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