title: Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment.
abstract: Antibody-immobilized thermoresponsive poly(N-isopropylacrylamide-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 carryover 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.
authors: Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University (TWIns), 8-1 Kawadacho, Shinjuku-ku, Tokyo, 162-8666, Japan.
authors: Jun
authors: J
authors: Kobayashi
authors: Masaki
authors: M
authors: Hayashi
authors: Takahiro
authors: T
authors: Ohno
authors: Masanori
authors: M
authors: Nishi
authors: Yoshinori
authors: Y
authors: Arisaka
authors: Yoshinori
authors: Y
authors: Matsubara
authors: Hiroshi
authors: H
authors: Kakidachi
authors: Yoshikatsu
authors: Y
authors: Akiyama
authors: Masayuki
authors: M
authors: Yamato
authors: Akihiro
authors: A
authors: Horii
authors: Teruo
authors: T
authors: Okano
mesh_terms: D000906:Antibodies / Q000737:chemistry*; D002448:Cell Adhesion; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry*; D013499:Surface Properties; D018800:Thy-1 Antigens / Q000096:biosynthesis*
chemical_list: D000906:Antibodies; D055767:Immobilized Proteins; D018800:Thy-1 Antigens
keywords: adipose tissue-derived cell; antibody; cell sheet; poly(N-isopropylacrylamide); thermoresponsive cell culture surface