The primary study of optimization parameters of ultrasonic microbubbles delivery hAng -1 gene into 2 9 3 T cells in vitro
10.3760/cma.j.issn.1004-4477.2009.12.021
- VernacularTitle:超声微泡介导体外转染hAng-1基因最佳条件的初步探讨
- Author:
Qing ZHOU
;
Qian CHEN
;
Xiao WANG
;
Jinling CHEN
;
Ruiqiang GUO
- Publication Type:Journal Article
- Keywords:
Ultrasonography;
Microbubbles;
Transfection
- From:
Chinese Journal of Ultrasonography
2009;18(12):1076-1079
- CountryChina
- Language:Chinese
-
Abstract:
Objective To certificate the effects on transfection ratio and cells viability of ultrasound (US) acoustic intensity, radiation duration, microbubbles concentration and DNA concentration in delivery human angiopioetin-1 gene (hAng-1) into 293T cells by SonoVue microbubbles and decide the optimal transfection parameters. Methods Mix 293T cells and SonoVue microbubbles linked with eGFP-C_3-hAng-1 in a different way, detect the gene transfection ratio and cells viability under the various US intensity, radiation duration, microbubbles and DNA concentrations. Results The gene expression would be increased if enhanced the intenstiy of US,radiation time,microbubbles and DNA concentrations,and the cells viability would be kept more than 90% ( P <0. 01). Whereas,if the US intensity increased over 1. 5 W/cm~2 ,the duration over 30 s and microbubbles and DNA concentrations over 20% and 15 mg/L respectively,the gene expression would not increase significantly ( P > 0. 05),whereas coupled with obviously decreased cells viability( P <0. 01). Conclusions The optimal conditions of deliver hAng-1 gene into 293T cells by SonoVue microbubbles was mixing cells and microbubbles in a cell wall-sticky way,US intensity was 1. 5 W/cm~2, duration 30 s,20% microbubbles and 15 mg/L DNA concentration.
