title: Culture on a fragmin/protamine-coated plate suppresses the collagen type IαI and TGF-β1 mRNA expression of rat hepatic stellate RI-T cells.
abstract: Hepatic stellate cells (HSCs) intracellularly preserve vitamin A in the normal liver. When the liver is damaged, HSCs transform into myofibroblast-like cells, and then proliferate and increase their expression of collagen. Cultured on a plastic plate, HSCs spontaneously activate. To maintain HSCs in a quiescent state with low expression of collagen, coating methods with extracellular matrixes (ECMs) such as Matrigel-coating or laminin-rich coating are commonly used for HSC cultivation. Kishimoto et al. [14] reported that Fragmin®/protamine microparticles (F/P-MPs) have the ability to absorb heparin-binding cytokines like ECMs. Therefore, we examined whether the cultivation on an F/P-MPs-coated plate maintains the quiescent state of RI-T cells (derived from rat HSCs) including the suppression of collagen expression. We found that the mRNA levels of collagen type IαI and TGF-β1 in RI-T cells were significantly suppressed in the cultivation on F/P-MPs-coated plates compared to cultures on noncoated and Matrigel-coated plates. We conclude that the F/P-MPs coating method is useful for maintaining with low expressions of collagen IαI and TGF-β 1 mRNA levels in HSCs.
authors.affiliation: Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1-17-71 Fuchinobe, Chuoh-ku, Sagamihara 252-5201, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Sekiguchi
authors.forename: Natsuko
authors.initials: N
authors.lastname: Hemmi
authors.forename: Takehiro
authors.initials: T
authors.lastname: Maki
authors.forename: Aisa
authors.initials: A
authors.lastname: Ozawa
authors.forename: Erina
authors.initials: E
authors.lastname: Kadowaki
authors.forename: Junichi
authors.initials: J
authors.lastname: Kamiie
authors.forename: Masako
authors.initials: M
authors.lastname: Yamamoto
authors.forename: Kazuyoshi
authors.initials: K
authors.lastname: Arishima
authors.forename: Motoharu
authors.initials: M
authors.lastname: Sakaue
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D055252:Cell-Derived Microparticles / Q000378:metabolism; D003094:Collagen; D024042:Collagen Type I / Q000378:metabolism*; D017931:DNA Primers / Q000235:genetics; D017985:Dalteparin / Q000494:pharmacology*; D004338:Drug Combinations; D005786:Gene Expression Regulation / Q000187:drug effects*; D055166:Hepatic Stellate Cells / Q000378:metabolism*; D007797:Laminin; D011479:Protamines / Q000494:pharmacology*; D011509:Proteoglycans; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D053773:Transforming Growth Factor beta1 / Q000378:metabolism*
chemical_list: D024042:Collagen Type I; D017931:DNA Primers; D004338:Drug Combinations; D007797:Laminin; D011479:Protamines; D011509:Proteoglycans; D053773:Transforming Growth Factor beta1; C059485:matrigel; D003094:Collagen; D017985:Dalteparin