Culture on a Fragmin/protamine-Coated Plate Suppresses the Collagen Type IαI and TGF-β1 mRNA Expression of Rat Hepatic Stellate RI-T Cells

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  • SEKIGUCHI Hitomi
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan
  • HEMMI Natsuko
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan
  • MAKI Takehiro
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan
  • OZAWA Aisa
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan Department of Veterinary Anatomy, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1–1–1 Yayoi, Bunkyo-ku, Tokyo 113–8657, Japan
  • KADOWAKI Erina
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan
  • KAMIIE Junichi
    Laboratory of Veterinary Pathology, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan
  • YAMAMOTO Masako
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan
  • ARISHIMA Kazuyoshi
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan
  • SAKAUE Motoharu
    Department of Anatomy II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Chuoh-ku, Sagamihara 252–5201, Japan

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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.

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