Switching of cell growth/detachment on heparin-functionalized thermoresponsive surface for rapid cell sheet fabrication and manipulation

Yoshinori Arisaka, Jun Kobayashi, Masayuki Yamato, Yoshikatsu Akiyama, Teruo Okano

    Research output: Contribution to journalArticle

    44 Citations (Scopus)

    Abstract

    Heparin-functionalized poly(N-isopropylacrylamide- co-2-carboxyisopropylacrylamide) [P(IPAAm- co-CIPAAm)] grafted surface was designed for the switching of cell growth/detachment, achieved by the regulation of affinity binding between basic fibroblast growth factor (bFGF) and immobilized heparin through the temperature-dependent conformational change of grafted P(IPAAm- co-CIPAAm) chains. At 37 °C, bFGF-bound heparin-thermoresponsive surfaces were able to hold the two- to three-fold number of mouse fibroblast (NIH/3T3) cells than both bFGF-physisorbed surface and PIPAAm surface with soluble bFGF after a 3-day cultivation. Bound bFGF via heparin on shrunken grafted P(IPAAm- co-CIPAAm) chains at 37 °C was able to reinforce the formation and stabilization of bFGF-FGF receptor complex, although the activity of physisorbed bFGF on PIPAAm-grafted surfaces was decreased by non-specific and randomly oriented adsorption. At 20 °C, the cultured NIH/3T3 cell sheet with bFGF detached from heparin-functionalized thermoresponsive surface. The release of bFGF from the surfaces was induced by reducing the affinity binding between bFGF and immobilized-heparin due to increasing the mobility of the swollen grafted P(IPAAm- co-CIPAAm) chains. Therefore, heparin-functionalized thermoresponsive surface was able to enhance cell proliferation, and confluent cells detached themselves as a contiguous cell sheet due to switching cell growth by changing temperature. A cell culture system using this surface is useful for rapid cell sheet fabrication and manipulation.

    Original languageEnglish
    Pages (from-to)4214-4222
    Number of pages9
    JournalBiomaterials
    Volume34
    Issue number17
    DOIs
    Publication statusPublished - 2013 Jun

    Fingerprint

    Cell growth
    Fibroblast Growth Factor 2
    Fibroblasts
    Heparin
    Fabrication
    Growth
    Fibroblast Growth Factor Receptors
    NIH 3T3 Cells
    Intercellular Signaling Peptides and Proteins
    Temperature
    Cell proliferation
    Cell culture
    Adsorption
    Stabilization
    Cell Culture Techniques
    Cell Proliferation
    2-carboxyisopropylacrylamide

    Keywords

    • Affinity
    • Basic fibroblast growth factor
    • Cell sheet
    • Heparin
    • Poly(N-isopropylacrylamide)
    • Thermoresponsive cell culture surface

    ASJC Scopus subject areas

    • Biomaterials
    • Bioengineering
    • Ceramics and Composites
    • Mechanics of Materials
    • Biophysics

    Cite this

    Switching of cell growth/detachment on heparin-functionalized thermoresponsive surface for rapid cell sheet fabrication and manipulation. / Arisaka, Yoshinori; Kobayashi, Jun; Yamato, Masayuki; Akiyama, Yoshikatsu; Okano, Teruo.

    In: Biomaterials, Vol. 34, No. 17, 06.2013, p. 4214-4222.

    Research output: Contribution to journalArticle

    Arisaka, Yoshinori ; Kobayashi, Jun ; Yamato, Masayuki ; Akiyama, Yoshikatsu ; Okano, Teruo. / Switching of cell growth/detachment on heparin-functionalized thermoresponsive surface for rapid cell sheet fabrication and manipulation. In: Biomaterials. 2013 ; Vol. 34, No. 17. pp. 4214-4222.
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    AU - Akiyama, Yoshikatsu

    AU - Okano, Teruo

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    AB - Heparin-functionalized poly(N-isopropylacrylamide- co-2-carboxyisopropylacrylamide) [P(IPAAm- co-CIPAAm)] grafted surface was designed for the switching of cell growth/detachment, achieved by the regulation of affinity binding between basic fibroblast growth factor (bFGF) and immobilized heparin through the temperature-dependent conformational change of grafted P(IPAAm- co-CIPAAm) chains. At 37 °C, bFGF-bound heparin-thermoresponsive surfaces were able to hold the two- to three-fold number of mouse fibroblast (NIH/3T3) cells than both bFGF-physisorbed surface and PIPAAm surface with soluble bFGF after a 3-day cultivation. Bound bFGF via heparin on shrunken grafted P(IPAAm- co-CIPAAm) chains at 37 °C was able to reinforce the formation and stabilization of bFGF-FGF receptor complex, although the activity of physisorbed bFGF on PIPAAm-grafted surfaces was decreased by non-specific and randomly oriented adsorption. At 20 °C, the cultured NIH/3T3 cell sheet with bFGF detached from heparin-functionalized thermoresponsive surface. The release of bFGF from the surfaces was induced by reducing the affinity binding between bFGF and immobilized-heparin due to increasing the mobility of the swollen grafted P(IPAAm- co-CIPAAm) chains. Therefore, heparin-functionalized thermoresponsive surface was able to enhance cell proliferation, and confluent cells detached themselves as a contiguous cell sheet due to switching cell growth by changing temperature. A cell culture system using this surface is useful for rapid cell sheet fabrication and manipulation.

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