title: Optimization of the transductional efficiency of lentiviral vectors: effect of sera and polycations.
abstract: Lentiviral vectors are widely used as effective gene-delivery vehicles. Optimization of the conditions for efficient lentiviral transduction is of a high importance for a variety of research applications. Presence of positively charged polycations reduces the electrostatic repulsion forces between a negatively charged cell and an approaching enveloped lentiviral particle resulting in an increase in the transduction efficiency. Although a variety of polycations are commonly used to enhance the transduction with retroviruses, the relative effect of various types of polycations on the efficiency of transduction and on the potential bias in the determination of titer of lentiviral vectors is not fully understood. Here, we present data suggesting that DEAE-dextran provides superior results in enhancing lentiviral transduction of most tested cell lines and primary cell cultures. Specific type and source of serum affects the efficiency of transduction of target cell populations. Non-specific binding of enhanced green fluorescent protein (EGFP)-containing membrane aggregates in the presence of DEAE-dextran does not significantly affect the determination of the titer of EGFP-expressing lentiviral vectors. In conclusion, various polycations and types of sera should be tested when optimizing lentiviral transduction of target cell populations.
authors.affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, AL 35294-2182, USA.
authors.forename: Warren
authors.initials: W
authors.lastname: Denning
authors.forename: Suvendu
authors.initials: S
authors.lastname: Das
authors.forename: Siqi
authors.initials: S
authors.lastname: Guo
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.forename: John C
authors.initials: JC
authors.lastname: Kappes
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Hel
mesh_terms: D002460:Cell Line; D003637:DEAE-Dextran / Q000032:analysis / Q000737:chemistry; D005434:Flow Cytometry; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D011073:Polyamines / Q000737:chemistry*; D000071228:Polyelectrolytes; D014161:Transduction, Genetic
chemical_list: D003637:DEAE-Dextran; D049452:Green Fluorescent Proteins; D011073:Polyamines; D000071228:Polyelectrolytes; C485184:enhanced green fluorescent protein