The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets

Yukiko Tsuda, Akihiko Kikuchi, Masayuki Yamato, Aiko Nakao, Yasuhisa Sakurai, Mitsuo Umezu, Teruo Okano

    Research output: Contribution to journalArticle

    151 Citations (Scopus)

    Abstract

    Heterotypic cell interactions are critical to achieve and maintain specific functions in many tissues and organs. We have focused on patterned structure surfaces to enable co-culture of heterotypic cells and recovery of patterned co-cultured cell sheets for applications in tissue engineering. Thermoresponsive polymers exhibiting different transition temperatures in water comprise both poly(N-isopropylacrylamide) (PIPAAm) and n-butyl methacrylate (BMA) co-grafted as side chains to PIPAAm main chains. These copolymers were surface-grafted in patterns to obtain patterned dual thermoresponsive cell culture surfaces using electron beam polymerisation method and porous metal masks. On patterned surfaces, site-selective adhesion on and growth of rat primary hepatocytes (HCs) and bovine carotid endothelial cells (ECs) allowed patterned co-culture, exploiting hydrophobic/hydrophilic surface chemistry regulated by culture temperature as the sole variable. At 27°C, seeded HCs adhered exclusively onto hydrophobic, dehydrated P(IPAAm-BMA) co-grafted domains (1-mm∅ area), but not onto neighbouring hydrated PIPAAm domains. Sequentially seeded ECs then adhered exclusively to hydrophobised PIPAAm domains upon increasing culture temperature to 37°C, achieving patterned co-cultures. Reducing culture temperature to 20°C promoted hydration of both polymer-grafted domains, permitting release of the co-cultured, patterned cell monolayers as continuous cell sheets with heterotypic cell interactions. Recovered co-cultured cell sheets can be manipulated, moved and sandwiched with other structures, providing new useful constructs both for basic cell biology research and preparation of tissue-mimicking multi-layer materials through overlaying co-cultured cell sheets.

    Original languageEnglish
    Pages (from-to)1885-1893
    Number of pages9
    JournalBiomaterials
    Volume26
    Issue number14
    DOIs
    Publication statusPublished - 2005 May

    Fingerprint

    Cultured Cells
    Coculture Techniques
    Endothelial cells
    Cells
    Cell culture
    Recovery
    Cell Communication
    Cytology
    Tissue
    Temperature
    Hepatocytes
    Polymers
    Endothelial Cells
    Surface chemistry
    Tissue engineering
    Surface structure
    Hydration
    Superconducting transition temperature
    Transition Temperature
    Rats

    Keywords

    • Cell culture
    • Cell sheet
    • Co-culture
    • n-Butyl methacrylate
    • Poly(N-isopropylacrylamide)
    • Surface patterning
    • Thermoresponsive surfaces

    ASJC Scopus subject areas

    • Biotechnology
    • Bioengineering
    • Biomedical Engineering

    Cite this

    The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets. / Tsuda, Yukiko; Kikuchi, Akihiko; Yamato, Masayuki; Nakao, Aiko; Sakurai, Yasuhisa; Umezu, Mitsuo; Okano, Teruo.

    In: Biomaterials, Vol. 26, No. 14, 05.2005, p. 1885-1893.

    Research output: Contribution to journalArticle

    Tsuda, Yukiko ; Kikuchi, Akihiko ; Yamato, Masayuki ; Nakao, Aiko ; Sakurai, Yasuhisa ; Umezu, Mitsuo ; Okano, Teruo. / The use of patterned dual thermoresponsive surfaces for the collective recovery as co-cultured cell sheets. In: Biomaterials. 2005 ; Vol. 26, No. 14. pp. 1885-1893.
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