Transfection of a mouse dendritic cell line by plasmid DNA-loaded PLGA microparticles in vitro.

摘要

Targeting of DC for DNA vaccination may be achieved by DNA-loaded poly(lactide-co-glycolide) (PLGA) biodegradable microparticles, since DC efficiently capture these microparticles in vitro and in vivo. DNA was encapsulated in PLGA microparticles by spray-drying. Various additives were tested and process parameters adjusted in order to prevent degradation of the DNA during encapsulation. The highest degree of supercoiled DNA was maintained by adding a strong buffering agent, such as PBS or NaHCO(3), whereas the cryoprotective lactose did not show a significant protective effect. DNA-containing PLGA microparticles were administered to a mouse DC line. Transfection efficacy was compared with commonly employed cationic transfectants and was visually assessed by green fluorescent protein expression. Transfection rate was very low in DC for all microparticle formulations and was comparable with commonly used cationic transfectants. It is concluded that the transfection of DC using PLGA microparticles is feasible, but efforts need to be undertaken to improve transfection efficiency in vitro, which may in addition lead to improved immune responses in vivo.