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

D. FORCATO; F. ALUSTIZA; A. FILI; J. LAZARO MARTINEZ; S. BONGIOVANNI ABEL; F. OLMOS NICOTRA; A. ALESSIO; N. RODRIGUEZ; C. A. BARBERO; P. BOSCH

CYTOTECHNOLOGY (DORDRECHT)

SPRINGER

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 polyethylenimine (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 transfection efficiency on bovine fetal fibroblasts. We found that transfection efficiency was affected mainly by polymer/pDNA ratio and DNA concentration and in less extent by PEI MW. In addition, preincubation of PEI nanoparticles with fetal bovine serum greatly increased percentage of cells expressing the transgene (up to 82%) while significantly decreased the polymer cytotoxic effect. In conclusion, our results indicate that fully deacylated PEI of 87 and 217 KDa are useful DNA vehicles for non-viral transfection of primary cultures of bovine fetal fibroblast.