Electroporation optimization to deliver plasmid DNA into dental follicle cells
| Autor: | Shaomian Yao, Gary E. Wise, Dawen Liu, Samir Rana |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Dental follicle
biology Electroporation lac operon Dental Sac DNA General Medicine Transfection Applied Microbiology and Biotechnology Molecular biology Article Rats Cell biology chemistry.chemical_compound Plasmid chemistry biology.protein Animals Molecular Medicine Cattle Bovine serum albumin Fetal bovine serum Plasmids |
| Zdroj: | Biotechnology Journal. 4:1488-1496 |
| ISSN: | 1860-7314 1860-6768 |
| DOI: | 10.1002/biot.200900039 |
| Popis: | Electroporation DNA transfer is a simple and versatile approach to deliver genes. To develop an optimal electroporation protocol to deliver DNA into cells, we conducted square wave electroporation experiments with using rat dental follicle cells as follows: 1) the cells were electroporated at different electric field strengths with lac Z plasmid; 2) plasmid concentrations were tested to determine the optimal doses; 3) various concentrations of bovine serum albumin or fetal bovine serum were added to the pulsing buffer; and, 4) the pulsing durations were studied to determine the optimal duration. These experiments indicated that the optimal electroporation electric field strength was 375 V/cm, and that plasmid concentrations greater than 0.18 μg/μl were required to achieve high transfection efficiency. BSA or FBS 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 to 120 milliseconds (ms) at 375 V/cm. Thus, an improved electroporation protocol was established by optimizing the above parameters. In turn, this electroporation protocol can be used to deliver DNA into dental follicle cells to study the roles of candidate genes in regulating tooth eruption. |
| Databáze: | OpenAIRE |
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