S-phase synchronized CHO cells show elevated transfection efficiency and expression using CaPi

Abstract

Various methods exist to transfect mammalian cells in culture. It is generally accepted that individual methods have to be optimized for each of the cell lines or cell types used. Despitethe use of optimized protocols, significant day-to-day variationsin transfection efficiency regularly occur. We postulate that the`status' of cell populations prior to transfection is involved insuch variability. This study evaluates standardized transfectionsdone at different phases of the cell cycle. Cell synchronizationwas achieved using mimosine. Transfection efficiency was monitored by fluorescence quantification of GFP (Green Fluorescent Protein). We show that transfection using the calcium-phosphate-DNA co-precipitation method, at differentphases of the cell cycle, yields variable expression levels of GFP. Highest GFP expression levels were seen when transfecting cell populations with a dominant representation of S-phase-cells.