Electroporation optimization to deliver plasmid DNA into dental follicle cells.
Electroporation is a simple and versatile approach for DNA transfer but needs to be optimized for specific cells. We conducted square wave electroporation experiments for rat dental follicle cells under various conditions. These experiments indicated that the optimal electroporation electric field strength was 375 V/cm, and that plasmid concentrations greater than 0.18 microg/microL were required to achieve high transfection efficiency. BSA or fetal bovine serum in the pulsing buffer significantly improved cell survival and increased the number of transfected cells. The optimal pulsing duration was in the range of 45-120 ms at 375 V/cm. This electroporation protocol can be used to deliver DNA into dental follicle cells to study the roles of candidate genes in regulating tooth eruption. This is the first report showing the transfection of dental follicle cells using electroporation. The parameters determined in this study are likely to be applied to transfection of other fibroblast cells.
Yao
Shaomian
S
Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803, USA.
Rana
Samir
S
Liu
Dawen
D
Wise
Gary E
GE
D000818:Animals; D002417:Cattle; D004247:DNA / Q000235:genetics*; D003795:Dental Sac / Q000166:cytology* / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D010957:Plasmids / Q000235:genetics*; D051381:Rats
D004247:DNA