Using carbon nanotubes to deliver genes to hard-to-transfect mammalian primary fibroblast cells

Abstract

The insertion of DNA into primary mammalian cells is an important step towards gene therapy and production of transgenic animals. Recently, carbon nanotubes have been explored as an efficient novel non-viral system for delivering genes to cells due to their unique structure and properties. However, their potential for transfecting primary bovine cells in non-toxic concentrations has not been tested. The purpose of this study was to evaluate the cytotoxicity of carboxylic acid-functionalized multiwalled carbon nanotubes (COOH-MWCNTs) and their use to deliver plasmid DNA encoding the gene of green fluorescent protein to bovine primary fibroblast cells. Flow-cytometry cell viability results have shown the non-toxic nature of COOH-MWCNTs at low concentrations. The frequency shifts in Raman spectroscopy showed that the plasmid DNA connects to the nanomaterial. Fluorescence imaging, flow-cytometry and PCR analysis confirmed that the COOH-MWCNT nanovectors delivered pDNA into primary fibroblast cells successfully. The results show that COOH-MWCNTs can be attractive alternatives for delivery of DNA into hard-to-transfect primary bovine cells.