METHOD FOR 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 w ell on a substrate conjugated with type I collagen and the polymer. Us e 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 detachm ent of the cells. At each ratio tested, cell attachment was greater wi th type IV collagen than with type I collagen. When human cancer cells obtained from surgical specimens were cultured on type IV collagen-po lymer substrate with a medium containing dextran sulfate (10 mug/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 treat ment with ethyleneglycol bis(2-aminoethylether) tetraacetic acid and l ow-temperature treatment (at about 25-degrees-C) without trypsin, and the cells continued to proliferate. These observations suggest that cu lture with a type IV collagen-polymer substrate and a culture medium c ontaining dextran sulfate is useful for establishment of human cancer cell lines.