Investigating cellular electroporation using planar membrane models and miniaturized devices


Uitert, Iris van (2010) Investigating cellular electroporation using planar membrane models and miniaturized devices. thesis.

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Abstract:This thesis focuses on increasing our understanding of the electroporation process. Electroporation is a technique employed to introduce foreign molecules into cells that can normally not pass the cell membrane. By applying a short but high electric field, pores appear in the membrane through which these molecules can enter the cell. This process is crucial for a number of biotechnological and medical applications (such as drug delivery, particle delivery and gene transfection). This technique is studied with model systems for the cell membrane (bilayer lipid membranes or BLMs) as well as in cells. The membrane models are here employed to study the effect of the membrane composition on the pore formation process. The composition is altered by the addition of different types of phospholipids, cholesterol and proteins and the complexity of the models is increased from binary via ternary to quaternary systems. The results show that membrane composition can have a large effect on the potential required to form pores in the membranes. In addition to this, a microfluidic system for BLM experimentation is developed. In this device, the BLMs are created vertically, allowing for combining electrical and optical measurements. This is a major advantage over a conventional system where only electrical analysis is feasible. The main purpose of this device is as a platform for protein studies for drug screening purposes. Lastly, a cell monolayer electroporation device is employed to verify the hypothesis from the previous results with BLMs that polarized cells become electroporated at a different applied potential than non-polarized cells. In this device, cells are grown onto a layer of hydrogel on top of an electrode substrate or onto a bare electrode substrate, to induce membrane polarization or not.
Item Type:Thesis
Electrical Engineering, Mathematics and Computer Science (EEMCS)
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