Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment

Jun Kobayashi, Masaki Hayashi, Takahiro Ohno, Masanori Nishi, Yoshinori Arisaka, Yoshinori Matsubara, Hiroshi Kakidachi, Yoshikatsu Akiyama, Masayuki Yamato, Akihiro Horii, Teruo Okano

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    Abstract

    Antibody-immobilized thermoresponsive poly(Nisopropylacrylamide- co-2-carboxyisopropylacrylamide) [poly (IPAAm-co-CIPAAm)]-grafted cell culture surfaces were designed to enhance both the initial adhesion of weakly adhering cells and the ability of cells to detach in response to low temperature through the regulation of affinity binding between immobilized antibodies and antigens on the cellular surface. Ty-82 cells and neonatal normal human dermal fibroblasts (NHDFs), which express CD90 on the cell surface, adhered to anti-CD90 antibody-immobilized thermoresponsive surfaces at 37°C, a condition at which the grafted thermoresponsive polymer chains shrank. Adherent Ty-82 cells were detached from the surfaces by lowering the temperature to 20°C and applying external forces, such as pipetting, whereas cultured NHDF sheets spontaneously detached themselves from the surface in response to reduced temperature alone. When the temperature was decreased to 20°C, the swelling of grafted thermoresponsive polymer chains weakened the affinity binding between immobilized antibody and antigen on the cells due to the increasing steric hindrance of the polymer chains around the antigen-recognition site of the immobilized antibodies. No contamination was detected on cells harvested from covalently immobilized antibodies on the culture surfaces by low-temperature treatment, whereas a carry over of the antibody and avidin from the avidin-biotin binding surface was observed. Furthermore, the initial adhesion of adipose tissue-derived cells, which adhere weakly to PIPAAm-grafted surfaces, was enhanced on the antibody -immobilized thermoresponsive surfaces.

    Original languageEnglish
    Pages (from-to)3883-3893
    Number of pages11
    JournalJournal of Biomedical Materials Research - Part A
    Volume102
    Issue number11
    DOIs
    Publication statusPublished - 2014 Nov 1

    Fingerprint

    Immobilized Antibodies
    Cell culture
    Antibodies
    Antigens
    Temperature
    Polymers
    Avidin
    Fibroblasts
    Adhesion
    Biotin
    Swelling
    Contamination
    Cells
    Tissue

    Keywords

    • Adipose tissue-derived cell
    • Antibody
    • Cell sheet
    • Poly(N-isopropylacrylamide)
    • Thermoresponsive cell culture surface

    ASJC Scopus subject areas

    • Biomedical Engineering
    • Biomaterials
    • Ceramics and Composites
    • Metals and Alloys

    Cite this

    Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment. / Kobayashi, Jun; Hayashi, Masaki; Ohno, Takahiro; Nishi, Masanori; Arisaka, Yoshinori; Matsubara, Yoshinori; Kakidachi, Hiroshi; Akiyama, Yoshikatsu; Yamato, Masayuki; Horii, Akihiro; Okano, Teruo.

    In: Journal of Biomedical Materials Research - Part A, Vol. 102, No. 11, 01.11.2014, p. 3883-3893.

    Research output: Contribution to journalArticle

    Kobayashi, J, Hayashi, M, Ohno, T, Nishi, M, Arisaka, Y, Matsubara, Y, Kakidachi, H, Akiyama, Y, Yamato, M, Horii, A & Okano, T 2014, 'Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment', Journal of Biomedical Materials Research - Part A, vol. 102, no. 11, pp. 3883-3893. https://doi.org/10.1002/jbm.a.35064
    Kobayashi, Jun ; Hayashi, Masaki ; Ohno, Takahiro ; Nishi, Masanori ; Arisaka, Yoshinori ; Matsubara, Yoshinori ; Kakidachi, Hiroshi ; Akiyama, Yoshikatsu ; Yamato, Masayuki ; Horii, Akihiro ; Okano, Teruo. / Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment. In: Journal of Biomedical Materials Research - Part A. 2014 ; Vol. 102, No. 11. pp. 3883-3893.
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    AU - Matsubara, Yoshinori

    AU - Kakidachi, Hiroshi

    AU - Akiyama, Yoshikatsu

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    AU - Horii, Akihiro

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