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Cell sonoporation


Current involved researchers (LabTAU): Corentin Cornu (PhD student), Jacqueline Ngo (Engineer), Claude Inserra (Ass. Prof.), Jean-Louis Mestas (Inserm researcher), Jean-Christophe Bera (Prof.)

Current involved collaborative researchers: Christelle Der Loughian (Ass. Prof., MATEIS, Lyon), Jean-Paul Rieu (Prof., ILM, Lyon), Wen-Shiang Chen (Ass. Prof., NTUH, Taiwan)

Past involved researchers: Pauline Labelle (Postdoc LMA, Marseille), Cyril Desjouy (Postdoc LMFA, Lyon), Manuela Fouqueray (ex-Postdoc)



Cell sonoporation in cavitation-controlled condition

The phenomenon of ultrasound enhancement of molecular uptake , known as sonoporation, is based on the temporary increase of cell membrane permeability and its poration allowing molecule transfer. Ultrasound then reveals many attributes of an ideal transfer system, as it is low-invasive, well-tolerated by the patients, cost-effective and can achieve site-specific transfer of energy within the body as ultrasound can be focused. Nevertheless, ultrasound transfection efficiencies of sonoporation are lower than those of other methods, these limitations being mainly due to the lack of fundamental understanding of the mechanisms underlying sonoporation. Even if the mechanisms leadgin to the cell membrane poration are still not clearly established, the interaction between cells, ultrasound and microbubbles appears to be the main candidate through the phenomenon of cavitation. Considering the chaotic behavior of cavitation initiation and leveling, and consequently the unpredictable biological effects, a good balance between transfection efficiency and cell toxicity could be achieved by controlling acoustic cavitation in real-time during sonication.

By sonicating different suspended or adherent cell lines, with numerous molecules or drugs, it has been shown that controlling cavitation process during sonication could significantly enhances sonoporation efficiencies and in all situation increase the reproducibility of the attained sonoporation rate.




 Related publications:

[1]  Transfection of cells in suspension by ultrasound cavitation. Reslan L, Mestas JL, Herveau S, Bera JC, Dumontet C. J Controlled Release, 142, 251-252, 2010.

[2]  Stabilizing in-vitro ultrasound-mediated gene transfection by regulating cavitation. Lo CW, Desjouy C, Chen SR, Lee JL, Inserra C, Bera JC, Chen WS. Ultrason. Sonochemistry, 21(2):833-839, 2014.

[3]  Sonoporation of adherent cells in regulated ultrasound cavitation. Muleki Seya P, Fouqueray M, Ngo J, Poizat A, Inserra C, Bera JC. Ultrasound in Medicine and Biology, 41(4):1008-1019, 2015.


 Towards mechanisms underlying cell sonoporation and its visualization

In order to visualize bubble-cell interaction and in the aim of getting information on sonoporation process, ultrasound setup are developed with the possibility of being mounted on microscope stage for real-time visualization during s onication experiment. A photograph of such setup is presented below, including a culture well located between two opposite piezoelectric transducers, and one hydrophone in contact with the culture well for implementing the real-time cavitation-controlled system. The potential of generating cavitation bubbles in the vicinity of the well's bottom has been confirmed by cavitating atop a supported fluorescent lipid bilayer membrane (see below). Near-surface cavitation cloud well appear both in transmission (natural light) and in flurescence.




 Related publications:

[1]  Jumping acoustic bubbles on lipid bilayers. Der Loughian C, Muleki Seya P, Pirat P, Inserra C, Bera JC, Rieu JP. Soft Matter, 11:3460-3469, 2015.

[2] Monitoring and control of inertial cavitation activity for enhancing ultrasound transfection: the SonInCaRe project. C. Inserra, P. Labelle, C. Der Loughian, J.L. Lee, M. Fouqueray, J. Ngo, A. Poizat, C. Desjouy, B. Munteanu, C.W. Lo, C. Vanbelle, J.P. Rieu, W.S. Chen, J.C. Bera. IRBM, 35(2): 94-99, 2014.