Methodologic study of direct preparation for chromosome analysis of human solid brain tumors

Human malignant glioma grown in athymic nude mice (NHG-1) and three freshly resected human solid gliomas were used in the study of factors influencing the direct preparation (DP) for chromosome analysis of human solid tumors. The results showed that: 1) the length of time after the blood supply was obstructed was a major factor in reducing the success rate of DP, i.e., a 2-hour delay resulted in a significantly lowered metaphase number and after 4 hours almost no metaphases could be seen; 2) preserving tumor cells at 4°C may prolong the time limit to about 4 hours; 3) culture medium (RPMI 1640 and Eagle MEM) and bovine calf serum concentration (0%, 10%, 20%, and 30%) did not influence the success rate significantly; 4) colchicine concentration (0.025 μg/mL, 0.05 μg/mL, 0.1 μg/mL) and time of treatment (30 min, 90 min, or 180 min) mainly affected the quality of chromosomes observed but had little effect on the quantity of metaphases that might be obtained. Based on these results, we had a success rate of more than 80% in 72 xenografts and 22 human brain tumors.