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Thin-film assembly of diethanolamine-based lipidic material as potential gene carrier in mouse embryonic neural stem cells.

Journal of biomedical materials research. Part A (2008-08-07)
Ken-Ichi Kusumoto, Satoko Yamashita, Takahiro Nagata, Takeshi Ido, Itaru Hamachi, Tetsuyuki Akao
ABSTRACT

Understanding of lipidic materials used for gene delivery system is essential for the effective design and development of potential applications in basic and therapeutic research. This study aimed to evaluate the biological activity of totally synthesized ditetradecylacetyldiethanolaminetrimethylammonium (TMA-C2-DEA-C14) as gene carriers for neural stem cells. The transfer abilities were estimated by expressing green fluorescent protein (GFP) in mouse embryonic neural stem cells. Here, we demonstrate that lipidic assembly of TMA-C2-DEA-C14, which was self-organized by incubation in water for a month at 25 degrees C, can provide an efficient gene delivery with low cytotoxicity ( approximately 40% of GFP-expressed neural stem cells). However, when dispersed by ultrasonication, TMA-C2-DEA-C14 showed low effect ( approximately 4%). Moreover, electron microscopic analysis showed that TMA-C2-DEA-C14 assembly is characterized by thin-film structures with polygonal shapes ( approximately 2.7 mum), and after association with DNA, their structures dramatically changes to form liposome complexes that can effectively deliver DNA into the cellular cytoplasm of neural stem cells. Thus, TMA-C2-DEA-C14 assembly identified in this study was determined to have an effective activity as gene carriers for primary neural stem cells. Our findings suggest that this approach can serve as a novel model for the development of lipidic materials on nonviral gene delivery system.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Diethanolamine hydrochloride, 98%
Sigma-Aldrich
Diethanolamine, ACS reagent, ≥98.5%
Sigma-Aldrich
Diethanolamine, reagent grade, ≥98.0%
Sigma-Aldrich
Diethanolamine, puriss. p.a., ACS reagent, ≥99.0% (GC)
Supelco
Diethanolamine, analytical standard
Sigma-Aldrich
Diethanolamine, BioUltra, ≥99.5% (GC)
Supelco
Diethanolamine, Pharmaceutical Secondary Standard; Certified Reference Material