File(s) not publicly available
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, NSMouse 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 MethodsVolume
158Pagination
56-63ISSN
0165-0270Department/School
Menzies Institute for Medical ResearchPublisher
Elsevier Science BvPlace of publication
Po Box 211, Amsterdam, Netherlands, 1000 AeRepository Status
- Restricted