Development of a confocal ultrasound device using an inertial cavitation control for transfection in-vitro

9th International Symposium on Cavitation

Mestas, J.L.^1,2,3,7 ; Chettab, K.^4,5,6 ; Roux, S.^4,5,6 ; Prieur, F.^1,2,3 ; Lafond, M.^1,2,3 ; Dumontet, C.^4,5,6 ; Lafon, C.^1,2,3

INSERM, U1032, LabTau, Lyon

F-69003, France^1

Université de Lyon, Lyon

F-69003, France^2

Université Lyon 1, Lyon

F-69003, France^3

Université de Lyon 1, Lyon

69000, France^4

INSERM U1052, Lyon

69008, France^5

CNRS UMR 5286, Lyon

69008, France^6

INSERM, U1032, LabTAU, 151 cours Albert Thomas, Lyon Cedex

69424, France^7

关键词: Flow cytometry analysis;    Green fluorescent protein;    Inertial cavitation;    Inertial cavitation activity;    Real-time feedback;    Transfection efficiency;    Ultrasound cavitation;    Ultrasound devices;   

Others  :  https://iopscience.iop.org/article/10.1088/1742-6596/656/1/012003/pdf
DOI  :  10.1088/1742-6596/656/1/012003

来源: IOP

【 摘 要 】

Sonoporation using low-frequency high-pressure ultrasound (US) is a non-viral approach for in vitro and in vivo gene delivery. We developed a new sonoporation device designed for spatial and temporal control of ultrasound cavitation. This device was evaluated for the in vitro transfection efficiency of a plasmid coding for Green Fluorescent Protein (peGFP- C1) in adherent and non-adherent cell lines. The frequency spectrum of the signal receive by a hydrophone is used to compute a cavitation index (CI) representative of the inertial cavitation activity. The influence of the CI on transfection efficiency, as well as reproducibility were determined. A real-time feedback loop control on CI was integrated in the process to regulate the cavitation level during sonoporation. In both adherent and non-adherent cell lines, the sonoporation device produced a highly efficient transfection of peGFP-C1 (40-80%), as determined by flow cytometry analysis of GFP expression, along with a low rate of mortality assessed by propidium iodide staining. Moreover, the sonoporation of non-adherent cell lines Jurkat and K562 was found to be equivalent to nucleofection in terms of efficiency and toxicity while these two cell lines were resistant to transfection with lipofection.