title: Growth arrest of vascular smooth muscle cells in suspension culture using low-acyl gellan gum.
abstract: The proliferation of vascular smooth muscle cells (SMCs) causes restenosis in biomaterial vascular grafts. The purposes of this study were to establish a suspension culture system for SMCs by using a novel substrate, low-acyl gellan gum (GG) and to maintain SMCs in a state of growth inhibition. When SMCs were cultured in suspension with GG, their proliferation was inhibited. Their viability was 70% at day 2, which was maintained at more than 50% until day 5. In contrast, the viability of cells cultured in suspension without GG was 5.6% at day 2. By cell cycle analysis, the ratio of SMCs in the S phase when cultured in suspension with GG was lower than when cultured on plastic plates. In SMCs cultured in suspension with GG, the ratio of phosphorylated retinoblastoma (Rb) protein to Rb protein was decreased and p27Kip1expression was unchanged in comparison with SMCs cultured on plastic plates. In addition, SMCs could be induced to proliferate again by changing the culture condition from suspension with GG to plastic plates. These results suggest that our established culturing method for SMCs is useful to maintain SMCs in a state of growth inhibition with high viability.
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8675, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Natori
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8675, Japan.
authors.forename: Masachika
authors.initials: M
authors.lastname: Fujiyoshi
authors.affiliation: Division of Pharmacy, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8677, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Uchida
authors.affiliation: Biological Research Laboratories, Nissan Chemical Industries Ltd., 1470 Shiraoka, Shiraoka, Saitama, 369-0305, Japan.
authors.forename: Natsuki
authors.initials: N
authors.lastname: Abe
authors.affiliation: Biological Research Laboratories, Nissan Chemical Industries Ltd., 1470 Shiraoka, Shiraoka, Saitama, 369-0305, Japan.
authors.forename: Tatsuro
authors.initials: T
authors.lastname: Kanaki
authors.affiliation: Department of Molecular Cell Biology, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8675, Japan.
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Fukumoto
authors.affiliation: Department of Clinical Pharmacy, Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8675, Japan. iishii@faculty.chiba-u.jp.|Division of Pharmacy, Chiba University Hospital, 1-8-1 Inohana, Chuo-ku, Chiba, Chiba, 260-8677, Japan. iishii@faculty.chiba-u.jp.
authors.forename: Itsuko
authors.initials: I
authors.lastname: Ishii
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D049109:Cell Proliferation / Q000187:drug effects*; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000096:biosynthesis; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000494:pharmacology*; D011817:Rabbits; D016160:Retinoblastoma Protein / Q000096:biosynthesis / Q000235:genetics; D013535:Suspensions / Q000737:chemistry / Q000494:pharmacology
chemical_list: D003470:Culture Media; D011135:Polysaccharides, Bacterial; D016160:Retinoblastoma Protein; D013535:Suspensions; D050760:Cyclin-Dependent Kinase Inhibitor p27; C048288:gellan gum
keywords: Low-acyl gellan gum; Polysaccharide; Vascular smooth muscle cells; p27Kip1