Competitive electroporation formulation for cell therapy
Open Access
- 10 June 2011
- journal article
- research article
- Published by Springer Science and Business Media LLC in Cancer Gene Therapy
- Vol. 18 (8), 579-586
- https://doi.org/10.1038/cgt.2011.27
Abstract
Established cell transfection via nucleofection relies on nucleofection buffers with unknown and proprietary makeup due to trade secrecy, inhibiting the possibility of using this otherwise effective method for developing cell therapy. We devised a three-step method for discovering an optimal formulation for the nucleofection of any cell line. These steps include the selection of the best nucleofection program and known buffer type, selection of the best polymer for boosting the transfection efficiency of the best buffer and the comparison with the optimal buffer from an established commercial vendor (Amaxa). Using this three-step selection system, competitive nucleofection formulations were discovered for multiple cell lines, which are equal to or surpass the efficiency of the Amaxa nucleofector solution in a variety of cells and cell lines, including primary adipose stem cells, muscle cells, tumor cells and immune cells. Through the use of scanning electron microscopy, we have revealed morphological changes, which predispose for the ability of these buffers to assist in transferring plasmid DNA into the nuclear space. Our formulation may greatly reduce the cost of electroporation study in laboratory and boosts the potential of application of electroporation-based cell therapies in clinical trials.Keywords
This publication has 47 references indexed in Scilit:
- Combined Pulse Electroporation – A Novel Strategy for Highly Efficient Transfection of Human and Mouse CellsPLOS ONE, 2010
- Adipose-derived Stem Cells as Therapeutic Delivery Vehicles of an Oncolytic Virus for GlioblastomaMolecular Therapy, 2010
- Human stem cell delivery for treatment of large segmental bone defectsProceedings of the National Academy of Sciences, 2010
- Nucleic Acids Electrotransfer-Based Gene Therapy (Electrogenetherapy): Past, Current, and FutureMolecular Biotechnology, 2009
- Molecular Engineering of Viral Gene Delivery VehiclesAnnual Review of Biomedical Engineering, 2008
- Mechanistic Analysis of Electroporation-Induced Cellular Uptake of MacromoleculesExperimental Biology and Medicine, 2008
- Adipose Tissue–Derived Human Mesenchymal Stem Cells Mediated Prodrug Cancer Gene TherapyCancer Research, 2007
- Gamma‐glutamylcysteine synthetase‐based selection strategy for gene therapy of chronic granulomatous disease and graft‐vs.‐host diseaseEuropean Journal of Haematology, 2007
- Nucleofection: A New Method for Cutaneous Gene Transfer?Journal of Biomedicine and Biotechnology, 2006
- Ultra-low sized cross-linked polyvinylpyrrolidone nanoparticles as non-viral vectors for in vivo gene deliveryBiomaterials, 2006