Transfection of bovine fetal fibroblast with poly(ethylenimine) (PEI) nanoparticles: effect of particle size and presence of fetal bovine serum on transgene delivery and cytotoxicity

DIEGO FORCATO; FEDERICO ALUSTIZA; JUAN MANUEL LÁZARO MARTÍNEZ; ANDREA FILLI; ALEJANDRO BONGIOVANNI; MARCELO NICOTRA; ANDRES ALESSIO; CÉSAR BARBERO; PABLO BOSCH

CYTOTECHNOLOGY (DORDRECHT)

SPRINGER

Lugar: Berlin; Año: 2017 vol. 69 p. 655 - 665

0920-9069

The development of efficient transfection protocols for livestock cells is crucial for implementation of cell-based transgenic methods to produce genetically modified animals. We synthetized fully deacylated linear 22, 87 and 217 KDa poly(ethylenimine) (PEI) nanoparticles and compared their transfection efficiency and cytotoxicity to commercial branched 25 KDa PEI and linear 58 KDa poly(allylamine) hydrochloride (PAH). We studied the effect of PEI size and presence of serum on polymer/pDNA polyplexes and found that transfection efficiency was affected chiefly by polymer/pDNA ratio and DNA concentration and in less extent by PEI MW. We found that preincubation of PEI nanoparticles with fetal bovine serum greatly increased transfection efficiency up to 82% of cells expressing the transgene while significantly decreasing the polymer cytotoxic effect. In conclusion, our results indicate that fully deacylated PEI of 22, 87 and 217 KDa are useful DNA vehicles for non-viral transfection of primary cultures of fetal bovine fibroblast.