Design and Testing of a Disposable Flow Cuvette for Continuous Electroporation of a Bioreactor’s Initial Algae Cultivation

Abstract

Electroporation is a technique applied both in biomedical and biotechnological fields which uses a high-voltage electric current to temporarily destabilize the plasma membrane of living cells, permitting the introduction of small molecules as well as nucleic acids into the cytosol. Besides viral and chemical transfections, this method is a common way to manipulate living cells. However, the majority of electroporation machines available on the market can only work using batch-based cuvettes treating only a few micrograms of cells. To transform cells in the order of several grams in the quickest possible way, it is necessary to use a continuous-flow method. In this work, we present the design, electric and fluid dynamics simulations, construction and testing of a flow cuvette that can adapt to standard electroporator systems. The flow cuvette connected with a peristaltic pump was able to successfully electroporate 20 mL of medium containing microalgae cells in less than 5 min. Microalgae Scenedesmus almeriensis cells were transfected with a fluorescent siRNA oligo as well as magnetically transformed by introducing magnetic nanoparticles in their cytoplasm. The flow cuvette presented here offers a valid tool for the high-throughput transformation/transfection/transfer of both prokaryotic and eukaryotic organisms, especially suitable for bioreactor cultivation and other industrial biotechnological contexts.

Keywords

• electroporation
• microalgae
• magnetic nanoparticles
• fluorescent siRNA oligo
• 3D printing