TRANSFECTION OF FETAL BOVINE FIBROBLAST WITH CATIONIC POLYMERS

FORCATO, DIEGO; ALUSTIZA, FABRISIO; FILI, ALEJANDRO; ALESSIO, ANA PAULA; OLMOS NICOTRA, MARÍA FLORENCIA; RODRIGUEZ, NANCY; BONGIOVANNI ABEL, S; LAZARO MARTINEZ JUAN; BARBERO, CESAR; BOSCH, PABLO

La Falda, Córdoba

Conferencia; XIX Annual Scientific Meeting- CORDOBA BIOLOGY SOCIETY; 2013

CORDOBA BIOLOGY SOCIETY

The objective was to evaluate transfection efficiency and cytotoxicity of two cationic polymers: polyalilamine (PAA) and 25 KDa polyethylenimine (PEI) in bovine fetal fibroblasts (BFF). Cells were seeded on 24-well plates in DMEM/10% SFB. Different PAA and PEI/plasmidic DNA (encoding GFP) ratios were studied (2:1, 1:1 w/w). Percentage of green fluorescent cells was determined by flow cytometry 48 h after transfection. Cell viability was determined by MTT assay. A higher proportion of GFP-expressing cells was detected at PEI:DNA of 2:1 compared to 1:1 (1 μg of DNA; 38.4±1.9% vs 13.5±1.4%). Doubling the amount of DNA did not improve transfection efficiency (38.4±1.9% vs 32.4±1.9%). PEI at 2 and 4 μg/well caused significant cytotoxicity (54.7 ± 3.4% and 18.5 ± 5.7%, respectively). Transfection efficiency of PAA at 0.5 and 1 μg/well was marginal (~3% of GFP-positive cells) and the cytotoxic effect was similar to that caused by PEI. Using the same polymer concentrations (2 μg per well), the percentage of fluorescent cells was higher in PEI transfected cultures (32.4±1.9%) than those observed in PAA treated cells (12.7±1.3%). These results indicate that PEI 25KDa outperforms PAA´s ability to deliver plasmidic DNA into BFF.