Optimization of KYSE-30 Esophagus Cancer Cell Line Transfection Using Lipofectamine 2000

There are several methods for delivery of vectors into eukaryotic cell lines. Transfection with liposomes is an easy and accessible way. Lipofectamine 2000 is a transfection reagent with liposome structure. Despite having a specific protocol, the volume of this reagent should be optimized for use in different cell lines. The aim of this study was to optimize transfection of KYSE-30 cell line with pEGFP-NI vector using Lipofectamine 2000.
The vector was purified by plasmid extraction kit. Transfection of the KYSE-30 cell line was done using Lipofectamine 2000 and different concentrations of the vector. Expression of green fluorescent proteins (GFP) was evaluated by fluorescence microscopy, and later analyzed with ImageJ software.
Optimized concentration of plasmid (5μg) and volume of Lipofectamine 2000 (6μl) were determined for KYSE-30 cell line. GFP plasmid transfection using the determined values showed more than 65% efficiency in the KYSE-30 cell line. The quantity of DNA per transfection and volume of reagent were identified as essential factors for a successful transfection.
This study shows that lipofection with lipofectamine 2000 is an efficient method of gene delivery into KYSE-30 cell line.