Effect Of Ultrasound-exposure On Sheep Fetal Fibroblasts Transfection Efficiency

Transfection technology refers to the import of exogenous genes like DNA; RNAinto eukaryotic cells, which is a common tool to study gene expression regulation,mutation analysis, etc. For ultrasound has a good directivity and ability to penetrate,some damages like cell wall rupture have been observed that caused by low frequency,low power ultrasound, which may change the permeability of cell membrane, and letthe intracellular materials release or extra cellular material in. This study investigatedthe influence of ultrasound on the foetus fibroblasts (SFF) cell transduction efficiencyconducted by lipofectin. SFF cells were subjected to ultrasound with differentfrequency and time, The target gene was detected by PCR and RT-PCR. G418wasused for culling the monoclonal cells with transfected cells and counting transfectionefficiency.Main Results:1. The transfection efficiency of SFF been exposure prior to transfection, thetransfection efficiency of SFF been exposure after transfection (exposure time is60s,under2.5MHz)2. The gene transfection efficiency of SFF cells mixed with their lipofectin andplasmids (exposure time is60s, under2.5MHz) is the best9.00±0.64）.3. Cell viability under different ultrasound exposure conditions, SFF cells ofexposure (exposure time is60s, under2.5MHz) was higher than (exposure time is60s, under3.5MHz). And when the exposure frequency is5.0MHz, exposure for30s,69s and120s, cell viability would be obviously effected.