Use of inert gas jets to measure the forces required for mechanical gene transfection

BioMedical Engineering OnLine

Guillaume Chouinard-Pelletier¹  Mathieu Leduc¹  David Guay¹  Sylvain Coulombe¹  Richard L Leask²  Elizabeth AV Jones³ 

[1] Chemical Engineering Department, McGill University, 3610 rue University, Montréal, H3A 2B2, Canada

[2] Montreal Heart Institute, Montréal, H1T 1C8, Canada

[3] Lady Davis Institute, Jewish General Hospital, McGill University, 3755 Cote Ste- Catherine Rd, Montreal, QC, H3T 1E2, Canada

关键词: Gene expression;    Naked plasmid DNA;    Non-viral gene therapy;    Transfection;   

Others  :  798015
DOI  :  10.1186/1475-925X-11-67

 received in 2012-06-26, accepted in 2012-09-03,  发布年份 2012

【 摘 要 】

Background

Transferring genes and drugs into cells is central to how we now study, identify and treat diseases. Several non-viral gene therapy methods that rely on the mechanical disruption of the plasma membrane have been proposed, but the success of these methods has been limited due to a lack of understanding of the mechanical parameters that lead to cell membrane permeability.

Methods

We use a simple jet of inert gas to induce local transfection of plasmid DNA both in vitro (HeLa cells) and in vivo (chicken chorioallantoic membrane). Five different capillary tube inner diameters and three different gases were used to treat the cells to understand the dependency of transfection efficiency on the dynamic parameters.

Results

The simple setup has the advantage of allowing us to calculate the forces acting on cells during transfection. We found permeabilization efficiency was related to the dynamic pressure of the jet. The range of dynamic pressures that led to transfection in HeLa cells was small (200 ± 20 Pa) above which cell stripping occurred. We determined that the temporary pores allow the passage of dextran up to 40 kDa and reclose in less than 5 seconds after treatment. The optimized parameters were also successfully tested in vivo using the chorioallantoic membrane of the chick embryo.

Conclusions

The results show that the number of cells transfected with the plasmid scales with the dynamic pressure of the jet. Our results show that mechanical methods have a very small window in which cells are permeabilized without injury (200 to 290 Pa). This simple apparatus helps define the forces needed for physical cell transfection methods.

【 授权许可】

   
2012 Chouinard-Pelletier et al.; licensee BioMed Central Ltd.

【 图 表 】

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