title: Novel approach for transient protein expression in primary cultures of human dental pulp-derived cells.
abstract: Transfection is a powerful method for investigating variable biological functions of desired genes. However, the efficiency of transfection into primary cultures of dental pulp-derived cells (DPDC) is low. Therefore, using a recombinant vaccinia virus (vTF7-3), which contains T7 RNA polymerase, we have established a transient protein expression system in DPDCs. In this study, we used the human polymeric immunoglobulin receptor (pIgR) cDNA as a model gene. pIgR expression by the vTF7-3 expression system was confirmed by flow cytometry analysis and Western blotting. Furthermore, exogenous pIgR protein localized at the cell surface in DPDCs and formed a secretory component (SC). This suggests that exogenous pIgR protein expressed by the vTF7-3 expression system acts like endogenous pIgR protein. These results indicate the applicability of the method for cells outgrown from dental pulp tissue. In addition, as protein expression could be detected shortly after transfection (approximately 5h), this experimental system has been used intensely for experiments examining very early steps in protein exocytosis.
authors.affiliation: Department of Endodontics, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-8310, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Suguro
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Mikami
authors.forename: Rieko
authors.initials: R
authors.lastname: Koshi
authors.forename: Bunnai
authors.initials: B
authors.lastname: Ogiso
authors.forename: Eri
authors.initials: E
authors.lastname: Watanabe
authors.forename: Nobukazu
authors.initials: N
authors.lastname: Watanabe
authors.forename: Masaki J
authors.initials: MJ
authors.lastname: Honda
authors.forename: Masatake
authors.initials: M
authors.lastname: Asano
authors.forename: Kazuo
authors.initials: K
authors.lastname: Komiyama
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus; D002478:Cells, Cultured; D006224:Cricetinae; D003782:Dental Pulp / Q000166:cytology* / Q000378:metabolism*; D005347:Fibroblasts; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D019056:Receptors, Polymeric Immunoglobulin / Q000737:chemistry* / Q000378:metabolism; D014162:Transfection; D014616:Vaccinia virus
chemical_list: D011506:Proteins; D019056:Receptors, Polymeric Immunoglobulin