Título:	Polyethyleneimine-mediated gene transfection in porcine fetal fibroblasts.
Autoria:	SOUZA, A. P. de BASTOS, A. P. A. FONSECA, F. M. da PANDOLFI, J. R. C. COSTAMILAN, C. A. da V. L. R. MARQUES, M. G.
Afiliação:	ANDRESSA PEREIRA DE SOUZA, UNIVERSIDADE DO ESTADO DE SANTA CATARINA; ANA PAULA ALMEIDA BASTOS, CNPSA; FRANCISCO NOE DA FONSECA, GGINT; JOSE RODRIGO CLAUDIO PANDOLFI, CNPSA; CARLOS ANDRÉ DA VEIGA LIMA ROSA COSTAMILAN, UNIVERSIDADE DO ESTADO DE SANTA CATARINA; MARIANA GROKE MARQUES, CNPSA.
Ano de publicação:	2024
Referência:	Animal Reproduction, v. 21, n. 4, e20240026, 2024.
Conteúdo:	Abstract: Polyethylenimine (PEI) has been explored as an efficient non-viral system for delivering genes to cells; however, there were no protocols for its use in porcine fetal fibroblasts (PFF). Therefore, we compared different concentrations of FITC-PEI (0.625, 1.25, 2.5, 5, 10, 20, 40, or 80 µg/mL) and incubation times (30 min, 1 h, or 2 h). It was observed that the incubation time did not affect the internalization of the PEI-FITC and that 30 min was sufficient to capture the complex. The concentrations higher than 10 µg/mL could reach many marked PFF (>90%). Then, two PEI concentrations were tested, 10 or 40 µg/mL, combined with an N/P of 2 with the pmhyGENIE-5 for 30 min. The percentage of PFF-GFP positive was similar between the PEI concentrations in the evaluation time points (24 h, 48 h, and 72 h). However, 40 µg/mL caused higher membrane damage rates. Thus, it can be concluded that concentrations between 10 – 80 µg/ml of PEI promote high incorporation rates, even in periods as short as 30 minutes. Furthermore, it can be stated that the transfection condition used in Polyplexes 1 (10 µg/mL of PEI and 37.5 µg/mL of pmhyGENIE5 for 30 min) efficiently produces genetically edited porcine fetal fibroblasts with low cell damage.
Thesagro:	Genética Animal Suinocultura Feto
NAL Thesaurus:	Animal genetics Swine
Palavras-chave:	Fibroblastos Transfecção genética Primary fibroblast Cationic polymer Non-viral vectors Gene delivery Nuclear transfer
Digital Object Identifier:	https://doi.org/10.1590/1984-3143-AR2024-0026
Tipo do material:	Artigo de periódico
Acesso:	openAccess
Aparece nas coleções:	Artigo em periódico indexado (CNPSA)