title: D-glucosamine promotes transfection efficiency during electroporation.
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.
authors.affiliation: Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
authors.forename: Kazunari
authors.initials: K
authors.lastname: Igawa
authors.affiliation: Department of Conservative Dentistry, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan.
authors.forename: Naoko
authors.initials: N
authors.lastname: Ohara
authors.affiliation: Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kawakubo
authors.affiliation: Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
authors.forename: Kouji
authors.initials: K
authors.lastname: Sugimoto
authors.affiliation: Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
authors.forename: Kajiro
authors.initials: K
authors.lastname: Yanagiguchi
authors.affiliation: Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Ikeda
authors.affiliation: Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
authors.forename: Shizuka
authors.identifier: 0000-0001-5346-5627
authors.initials: S
authors.lastname: Yamada
authors.affiliation: Department of Cariology, Nagasaki University School of Biomedical Sciences, Nagasaki 852-8588, Japan.
authors.forename: Yoshihiko
authors.identifier: 0000-0002-4256-3085
authors.initials: Y
authors.lastname: Hayashi
mesh_terms: D002021:Buffers; D002460:Cell Line; D004566:Electrodes; D018274:Electroporation / Q000379:methods*; D005944:Glucosamine / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D014162:Transfection
chemical_list: D002021:Buffers; D049452:Green Fluorescent Proteins; D005944:Glucosamine