Efficient establishment of Guangxi Bama mini-pig transgenic fibroblasts via Xfect polymer transfection

摘要

A variety of vectors have been developed to deliver DNA into porcine somatic cells, nevertheless, the concerns of bio-safety, operational complexity, high cytotoxicity and low transfection efficiency restrict their widespread applications in genetic modification of porcine somatic cells. Xfect polymer is a novel biodegradable transfection reagent has been employed to deliver exogenous genes into several types of eukaryotic cell with minimal cytotoxicity and superior transfection efficiency. This study was designed to examine whether Xfect polymer is suitable for genetically modifying porcine somatic cells. Transfection conditions including dosage of plasmid DNA and duration of polyplexes incubation were optimized. Then, optimized transfection conditions combining with G418 screening were used to establish Guangxi Bama mini-pig transgenic fibroblast cell lines. Intense expression of red fluorescence protein as a reporter gene in porcine transgenic fibroblasts was detected using a fluorescent microscopy. Furthermore, good cell proliferation and almost no morphological signs of cell stress were also observed. Therefore, our findings suggest Xfect polymer could effectively introduce exogenous genes into porcine somatic cells serves as an alternative transfection method.