Table 1.

Comparison of Different iCELLis Runs

Run Number	Fixed-Bed	Working Volume	Media before Transfection	Media PT	Type of Media Supply	DNA/cm2	Virus Produced	Endonuclease Treatment
Run 1	HC, 100 m2	65 L	DMEM, 10% FBS, L-glut.	DMEM, L-glut.	perfusion aiming 0.5 g/L glucose	300 ng	LV-GFP	after harvest
Nano 1	LC, 2.67 m2	800 mL	DMEM, 10% FBS, L-glut., P/S	DMEM, L-glut., P/S	perfusion aiming 0.5 g/L glucose	300 ng	LV-GFP	after harvest
Nano 2	LC, 2.67 m2	800 mL	DMEM, 10% FBS, L-glut., P/S	DMEM, L-glut., P/S	recirculation	300 ng	LV-GFP	after harvest
Nano 3	LC, 2.67 m2	800 mL	DMEM, 10% FBS, L-glut., P/S	DMEM, L-glut., P/S	perfusion aiming 0.5 g/L glucose	300 ng	LV-GFP	after harvest
Nano 4	LC, 2.67 m2	800 mL	DMEM, 10% FBS, L-glut., P/S	DMEM, L-glut., P/S, extras	perfusion aiming 0.5 g/L glucose	200 ng	LV-GFP	after harvest
Nano 5	LC, 2.67 m2	800 mL	DMEM, 10% FBS, L-glut., P/S	DMEM, L-glut., P/S, extras	perfusion aiming 0.5 g/L glucose	200 ng	LV-TK	after harvest
Run 2	LC, 333 m2	65 L	DMEM, 10% FBS, L-glut.	DMEM, L-glut., extras	perfusion aiming 0.5 g/L glucose	200 ng	LV-TK	during production in bioreactor and after harvest

In all runs, 7,000 cells/cm² were inoculated and pH was set to 7.2 before transfections and 7.0 after transfection. Abbreviations are as follows: HC, high compaction; LC, low compaction; PT, post-transfection; FBS, fetal bovine serum; L-glut., L-glutamine (4 mM); P/S, Penicillin-Streptomycin (50 μg/mL-50 U/mL); extras, NEAA + Na-pyruvate + CD-lipid supplement; LV, lentiviral vector; TK, thymidine kinase.