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|Title:||pH dependent high transfection efficiency of mouse neuroblastomas using TransFectin|
|Citation:||Chong, K.W.Y., Seet, S.J., Cheung, N.S., Lee, A.Y.-W., Koay, E.S.C. (2006). pH dependent high transfection efficiency of mouse neuroblastomas using TransFectin. Journal of Neuroscience Methods 158 (1) : 56-63. ScholarBank@NUS Repository. https://doi.org/10.1016/j.jneumeth.2006.05.017|
|Abstract:||Mouse neuroblastoma cell lines are often used in lieu of mouse primary neurons in ex vivo experiments, as they provide an easier platform for transfection, compared to the latter. A well-known inherent problem with this strategy is the relatively low transfection efficiency (15-30%) of mouse neuroblastoma cell lines such as neuro-2A and N1E-115. We were able to improve the transfection efficiency of these cell lines by using the cationic lipid reagent, TransFectin™ (Bio-Rad, Hercules, CA, USA) to optimise the transfection conditions. Our results, based on fluorescence intensity determinations and Western blotting for enhanced green fluorescence protein (EGFP) over-expression in neuro-2A, demonstrated that pH is a crucial factor in determining the transfection efficiency. Under pH-optimised transfection conditions, flow cytometric analysis revealed high EGFP transfection efficiencies of 76.4 ± 0.5 and 60.9 ± 0.6% for neuro-2A and N1E-115, respectively. Notably, the optimised TransFectin™-based transfection system did not result in any detectable cytotoxicity to the mouse neuroblastomas. The resultant optimised system is economical, easy to use and does not require any specialised equipment. © 2006 Elsevier B.V. All rights reserved.|
|Source Title:||Journal of Neuroscience Methods|
|Appears in Collections:||Staff Publications|
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