Liposomes-mediated chemosynthesis siRNA transfection to primary cardiomyocytes: Selection of an ideal concentration / 中国组织工程研究

ABSTRACT

BACKGROUND: siRNA transfection is a key step in RNA interference. The methods of cardiomyocytes transfection were most use of plasmids as vector to transfect long-chain shRNA. However, the processes were complex. It was a simple efficient and low-toxic method that liposomes-mediatad chernosynthesis siRNA transfection. It was useful for expanding RNA interference application.OBJECTIVE: To choice the optimal concentration of Iiposomes-mediated chemosynthesis siRNA transfection, and to discover a simple efficient RNA interference application.METHODS: CY3-Negative siRNA was mediated by lipid-besed agent siPORT~(TM) NeoFX~(TM) to transfect cardiomyocytes. A blank control group was set. After 24 hours, the transfection efficiency and apoptotic rate were evaluated by fluorescent microscope and flow cytometer to select an optimal concentration. Based the best concentration, siRNA PHB was transfected to cerdiomyocytes. 48 hours later, the expression of PHB was tested.RESULTS AND CONCLUSION: With the increased concentration of CY3-Negative siRNA, the number of cells emitted red fluorescence grew under fluorescence microscope, and the transfection efficiency was also increased (P＜0.05). The best concentration was 30 nmol/L (P＜0.05). There was no significant difference in apoptotic rate between transfected groups and the control group (P ＞ 0.05). The PHB expression of cardiomyocytes transfected siRNA PHB was dropped by 74.11% on average (P＜0.05). The results indicated that lipid based agent siPORT~(TM) NeoFX~(TM) was suitable to transfect chemosynthesis siRNA to cardiomyocytes, and the best transfection concentration of siRNA was 30 nmol/L.