Comparison of Various Transfection Methods in Human and Bovine Cultured Cells.
10.11620/IJOB.2014.39.4.177
- Author:
Longxun JIN
1
;
Daehwan KIM
;
Sangho ROH
Author Information
1. Cellular Reprogramming & Embryo Biotechnology Laboratory, Dental Research Institute and School of Dentistry, Seoul National University, Korea. sangho@snu.ac.kr
- Publication Type:Original Article
- Keywords:
transfection;
magnetofection;
non-liposomal transfection;
electroporation
- MeSH:
Cell Survival;
Cells, Cultured*;
Dental Pulp;
DNA;
Electroporation;
Fibroblasts;
Genes, vif;
Humans;
Indicators and Reagents;
Pluripotent Stem Cells;
Transfection*
- From:International Journal of Oral Biology
2014;39(4):177-185
- CountryRepublic of Korea
- Language:English
-
Abstract:
Transfection is a gene delivery tool that is a popular means of manipulating cellular properties, such as induced pluripotent stem cell (iPSC) generation by reprogramming factors (Yamanaka factors). However, the efficiency of transfection needs to be improved. In the present study, three transfection protocols - non-liposomal transfection (NLT), magnetofection and electroporation - were compared by analysis of their transfection efficiencies and cell viabilities using human dental pulp cells (hDPC) and bovine fetal fibroblasts (bFF) as cell sources. Enhanced green fluorescent protein gene was used as the delivery indicator. For magnetofection, Polymag reagent was administrated. NLT, FuGENE-HD and X-treme GENE 9 DNA transfection reagents were used for NLT. For electroporation, the Neon(TM) and NEPA21(TM) electroporators were tested. Neon(TM) electroporation showed highest transfection efficiency when compared with NLT, magnetofection, and NEPA21(TM) electroporation, with transfection efficiency of about 33% in hDPC and 50% in bFF, based on viable cell population in each cell type. These results suggest that transfection by Neon(TM) electroporation can be used to deliver foreign genes efficiently in human and bovine somatic cells.
