The effects of different ultrasound parameters and transfection conditions on the red fluorescent protein gene delivery and cell viability
- VernacularTitle:不同超声参数及转染方式对细胞活力和红色荧光蛋白基因传输的影响
- Author:
Zhiyi CHEN
;
Mingxing XIE
;
Xinfang WANG
;
Qing LU
- Publication Type:Journal Article
- Keywords:
Ultrasonography;
Gene transfer techniques;
Cell survival
- From:
Chinese Journal of Ultrasonography
2008;17(11):989-993
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate different ultrasound parameters and transfection conditions that would affect transfection rate of red fluorescent protein(RFP)and cell viability of cancer cells.Methods In this study,Hela cells were cultured using two different protocols:(A)24 h culture for complete adherence;(S)suspension.Subsequently,cells were transfected following different ultrasound exposure protocols[1.0W/cm2;duty cycle(DC):10%,20%and 50%;exposure 1min or 3 min].Gene transfection and cell viability were evaluated.Treatment parameters optimized in Hela cells were applied for delivery RFP in 4 other cell lines(HepG2,Ishikawa,MCF-7 and B16-F10).Results Cell injury were found to increase progressively with DC and exposure time in group A.Cell detachment was significantly accompanied by ultrasound exposure in adherent HeLa cells.Cells in group S were found more prone to be transfected than group A with the same ultrasound parameters,while the survival rate was not decreased apparently.The ideal ultrasound conditions were noted to be at 1.0 W/cm2 irradiated 3 min with 20%DC using suspended protocol,producing maximum efficiency[transfection=(28.04±2.27)%]in gene delivery with minimum cell toxicity[cell viability=(81.20±1.73)%].These experiments also revealed different response to ultrasound treatment,but for all tested cell lines,dead and transfected cells in the treated groups were significantly different from the non-irradiated groups.Conclusions Ultrasound parameters and transfection conditions have a great impact on the gene delivery and cell viability.Gene delivery of ultrasound-mediated microbubble enhance should be optimized to improve the efficiency.
