Simplified lentivirus vector production in protein-free media using polyethylenimine-mediated transfection

Evidence of an oncogenic role of aberrant TOX activation in cutaneous T-cell lymphoma

“…Lentiviral particles were generated by transfecting HEK 293T packaging cells using the polyethylenimine method as described previoiusly. 24 After 48 and 72 hours, the virus-containing medium (35 mL per construct) was filtered and concentrated in 200 mL of RPMI 1640 medium. CTCL cells (2 3 10 5 cells in 1 mL of media) were incubated with 5 mL of concentrated lentiviral particles for 24 hours.…”

Production of Lentiviral Vectors Encoding Recombinant Factor VIII Expression in Serum-Free Suspension Cultures

“…2B). A number of studies have reported optimal efficiency of transfection using DNA concentration in the range of 0.4-0.6 μg per 10 6 cells in PEI-mediated protocols (Sun et al 2008;Kuroda et al 2009). Besides, PEI is known to have a toxic effect (Godbey and Mikos 2001;Kunath et al 2003;Sun et al 2008) and the concentration of PEI: DNA complex (polyplex) may influence the efficiency of the transient transfection.…”

“…Currently, the most successful alternative for large scale LV production is based on transient transfection in serum-free suspension cultures using polyethylenimine (PEI) (Ansorge et al 2009;Schweizer and Merten 2010). PEI-mediated transfections have been reported to be more consistent, reproducible and productive (Pham et al 2006;Kuroda et al 2009;Toledo et al 2009). Moreover, PEI is chemically stable, inexpensive and has a broad pH range for transfection (Boussif et al 1995).…”

Efficient Large Volume Lentiviral Vector Production Using Flow Electroporation

“…Disadvantages of the calcium phosphate method include reagent pH sensitivity (Karolewski et al, 2003) and effectiveness only in adherent cell lines. Other transient transfection methods using lipofection (Felgner et al, 1987) or polyethylenimine have been reported (Kuroda et al, 2009;Segura et al, 2010). However, their use may add significant cost to large-scale, clinical productions when cGMP-grade transfection reagents must be used.…”