TY - JOUR
T1 - Surface design of antibody-immobilized thermoresponsive cell culture dishes for recovering intact cells by low-temperature treatment
AU - Kobayashi, Jun
AU - Hayashi, Masaki
AU - Ohno, Takahiro
AU - Nishi, Masanori
AU - Arisaka, Yoshinori
AU - Matsubara, Yoshinori
AU - Kakidachi, Hiroshi
AU - Akiyama, Yoshikatsu
AU - Yamato, Masayuki
AU - Horii, Akihiro
AU - Okano, Teruo
PY - 2014/11/1
Y1 - 2014/11/1
N2 - 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.
AB - 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.
KW - Adipose tissue-derived cell
KW - Antibody
KW - Cell sheet
KW - Poly(N-isopropylacrylamide)
KW - Thermoresponsive cell culture surface
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U2 - 10.1002/jbm.a.35064
DO - 10.1002/jbm.a.35064
M3 - Article
C2 - 24339415
AN - SCOPUS:84908063319
SN - 1549-3296
VL - 102
SP - 3883
EP - 3893
JO - Journal of Biomedical Materials Research - Part A
JF - Journal of Biomedical Materials Research - Part A
IS - 11
ER -