Method of Culture of Human Cancer Cells with a Thermoresponsive Polymer and Dextran Sulfate.

A method for cell culture with a thermoresponsive polymer and dextran sulfate was developed. In the culture of human cancer cell lines, the polymer had no cytotoxicity and the cells attached, spread, and grew well on a substrate conjugated with type I collagen and the polymer. Use of different collagen : polymer ratios in the primary culture of cells from human cancer which had serially been transplanted into nude mice showed that the ratio of 2 : 1 gave satisfactory attachment and detachment of the cells. At each ratio tested, cell attachment was greater with type IV collagen than with type I collagen. When human cancer cells obtained from surgical specimens were cultured on type IV collagen-polymer substrate with a medium containing dextran sulfate (10μg/ml), the growth of contaminating fibroblasts was suppressed and the cancer cells proliferated. During subculture, the cancer cells detached from the primary culture on the type IV collagen-polymer substrate by treatment with ethyleneglycol bis (2-aminoethylether) tetraacetic acid and low-temperature treatment (at about 25°C) without trypsin, and the cells continued to proliferate. These observations suggest that culture with a type IV collagen-polymer substrate and a culture medium containing dextran sulfate is useful for establishment of human cancer cell lines.