D-glucosamine promotes transfection efficiency during electroporation

Kazunari Igawa, Naoko Ohara, Atsushi Kawakubo, Kouji Sugimoto, Kajiro Yanagiguchi, Takeshi Ikeda, Shizuka Yamada, Yoshihiko Hayashi

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Abstract

D-Glucosamine is a useful medicament in various fields of medicine and dentistry. With respect to stability of the cell membrane, it has been reported that bradykinin-induced nociceptive responses are significantly suppressed by the direct application of D-glucosamine. Electroporation is usually used to effectively introduce foreign genes into tissue culture cells. Buffers for electroporation with or without D-glucosamine are used in experiments of transfection vectors. This is the first study to indirectly observe the stability and protection of the osteoblast membrane against both electric stress and gene uptake (the proton sponge hypothesis: osmotic rupture during endosomes prior to fusion with lysosomes) in electroporation with D-glucosamine application. The transfection efficiency was evaluated as the fluorescence intensity of the transfected green fluorescent protein (GFP) in the cultured cells (osteoblasts; NOS-1 cells). The transfection efficiency increased over 30% in the electroporation samples treated with D-glucosamine-supplemented buffer after one day. The membrane absorption of D-glucosamine is the primary mechanism of membrane stress induced by electric stress. This new function of D-glucosamine is useful and meaningful for developing more effective transformation procedures.

Original languageEnglish
Article number485867
JournalBioMed research international
Volume2014
DOIs
Publication statusPublished - 2014

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Cite this

Igawa, K., Ohara, N., Kawakubo, A., Sugimoto, K., Yanagiguchi, K., Ikeda, T., Yamada, S., & Hayashi, Y. (2014). D-glucosamine promotes transfection efficiency during electroporation. BioMed research international, 2014, [485867]. https://doi.org/10.1155/2014/485867