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  • thermoresponsive polymer  (2)
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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 53 (1997), S. 339-344 
    ISSN: 0006-3592
    Keywords: cell culture ; tissue engineering ; thermoresponsive polymer ; cell adhesion ; insulin conjugate ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: We developed a new biomaterial for use in cell culture. The biomaterial enabled protein-free cell culture and the recovery of viable cells by lowering the temperature without the aid of supplements. Insulin was immobilized and a thermoresponsive polymer was grafted onto a substrate. We investigated the effect of insulin coupling on the lower critical solution temperature (LCST) of the thermoresponsive polymer, poly(N-isopropylacrylamide-co-acrylic acid), using polymers that were ungrafted, or coupled with insulin. The insulin conjugates were precipitated from an aqueous solution at high temperatures, but they were soluble at low temperatures. The LCST was not significantly affected by the insulin coupling. The thermoresponsive polymer was grafted to glow-discharged polystyrene film and covalently conjugated with insulin. The surface wettability of the conjugate film was high at low temperatures and low at high temperatures. The amounts of immobilized insulin required to stimulate cell growth were 1-10% of the amount of free insulin required to produce the same effect. The maximal mitogenic effect of immobilized insulin was greater than that of free insulin. About half of the viable cells was detached from the film only by lowering the temperature. The recovered cells proliferated normally on new culture dishes. © 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 339-344, 1997.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Hoboken, NJ : Wiley-Blackwell
    Journal of Biomedical Materials Research 42 (1998), S. 38-44 
    ISSN: 0021-9304
    Keywords: pattern-grafting ; thermoresponsive polymer ; protein adsorption ; cell detachment ; cell culture engineering ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: A thermoresponsive copolymer, poly(N-isopropylacrylamide-co-acrylic acid), was coupled with azidoaniline. The azidophenyl-derivatized copolymer was grafted in a specific pattern on a polystyrene matrix by photolithography. The surface micropattern appeared and disappeared interchangeably, as observed under a phase-contrast microscope, by varying the temperature between 10°C and 37°C. The copolymer-grafted polystyrene surface was hydrophobic at 37°C and hydrophilic at 10°C. Albumin and fibronectin adsorption on the matrix was investigated using the fluorescent-labeling method. Fibronectin adsorbed onto both the grafted and nongrafted regions, while albumin adsorbed more onto the nongrafted regions than the grafted regions. Protein adsorption did not affect surface wettability. Mouse fibroblast STO cells were cultured on tissue culture plates pattern-grafted with the thermoresponsive copolymer. Fibronectin adsorption enhanced cell spreading, while albumin reduced it. When the temperature was lowered, the cells selectively detached from the surface areas grafted with the thermoresponsive copolymer when cultured in serum-free medium; the cells partially detached from these areas when cultured in serum-containing medium. The effect of serum proteins on cell detachment was similar to that caused by a mixture of albumin and fibronectin. Albumin adsorption did not affect the detachment of cells, while fibronectin adsorption inhibited it. The results of the present study indicate that a pattern-grafted, thermoresponsive, azidophenyl-derivatized copolymer can effectively facilitate selective cell detachment under some conditions such as serum-free culture or preadsorption of albumin. The pattern-grafting technique will be useful for qualitative microscopic comparison of surfaces prepared differently on one chip under the same conditions. © 1998 John Wiley & Sons, Inc. J. Biomed Mater Res, 42, 38-44, 1998.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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