Comparison of Various Transfection Methods in Human and Bovine Cultured Cells
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Transfection is a gene delivery tool that is a popular meansof manipulating cellular properties, such as inducedpluripotent stem cell (iPSC) generation by reprogrammingfactors (Yamanaka factors). However, the efficiency oftransfection needs to be improved. In the present study, threetransfection protocols - non-liposomal transfection (NLT),magnetofection and electroporation - were compared byanalysis of their transfection efficiencies and cell viabilitiesusing human dental pulp cells (hDPC) and bovine fetalfibroblasts (bFF) as cell sources. Enhanced green fluorescentprotein gene was used as the delivery indicator. Formagnetofection, Polymag reagent was administrated. NLT,FuGENE-HD and X-treme GENE 9 DNA transfectionreagents were used for NLT. For electroporation, the NeonTMand NEPA21TM electroporators were tested. NeonTMelectroporation showed highest transfection efficiency whencompared with NLT, magnetofection, and NEPA21TMelectroporation, with transfection efficiency of about 33% inhDPC and 50% in bFF, based on viable cell population ineach cell type. These results suggest that transfection byNeonTM electroporation can be used to deliver foreign genesefficiently in human and bovine somatic cells.
Abstract
Transfection is a gene delivery tool that is a popular meansof manipulating cellular properties, such as inducedpluripotent stem cell (iPSC) generation by reprogrammingfactors (Yamanaka factors). However, the efficiency oftransfection needs to be improved. In the present study, threetransfection protocols - non-liposomal transfection (NLT),magnetofection and electroporation - were compared byanalysis of their transfection efficiencies and cell viabilitiesusing human dental pulp cells (hDPC) and bovine fetalfibroblasts (bFF) as cell sources. Enhanced green fluorescentprotein gene was used as the delivery indicator. Formagnetofection, Polymag reagent was administrated. NLT,FuGENE-HD and X-treme GENE 9 DNA transfectionreagents were used for NLT. For electroporation, the NeonTMand NEPA21TM electroporators were tested. NeonTMelectroporation showed highest transfection efficiency whencompared with NLT, magnetofection, and NEPA21TMelectroporation, with transfection efficiency of about 33% inhDPC and 50% in bFF, based on viable cell population ineach cell type. These results suggest that transfection byNeonTM electroporation can be used to deliver foreign genesefficiently in human and bovine somatic cells.
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