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pH dependent high transfection efficiency of mouse neuroblastomas using TransFectin

journal contribution
posted on 2023-05-17, 01:43 authored by Chong, KW, Lee, AY, Koay, ES, Seet, SJ, Cheung, NS
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.

History

Publication title

Journal of Neuroscience Methods

Volume

158

Pagination

56-63

ISSN

0165-0270

Department/School

Menzies Institute for Medical Research

Publisher

Elsevier Science Bv

Place of publication

Po Box 211, Amsterdam, Netherlands, 1000 Ae

Repository Status

  • Restricted

Socio-economic Objectives

Clinical health not elsewhere classified

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