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Technigue and Method
Establishment of a human hepatocellular carcinoma cell line EH-H1 and its biological characteristics
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Abstract:
Objective:To establish a human hepatocellular carcinoma cell line EHH1 by primary culture and to investigate its biological characteristics. Methods: Tumor tissues obtained from patients treated in our hospital were made into singlecell suspension and were primary cultured, passaged in DMEM medium supplemented with 10% fetal bovine serum. The morphology of the cells was observed under light and electronic microscope; the cell growth curve was plotted. Radioimmunoassay was employed to examine the AFP and HBV expression. Cytogenetic study was performed with the cell line using Gbanding chromosome analysis. The tumorigenicity of the cell line was studied in nude mice by subcutaneous injection.Results: We have successfully established a novel hepatocellular carcinoma cell line EHH1, which could be passaged for over 80 generations with unchanged morphology and growth cycle (24 h). Radioimmunoassay results demonstrated that all EHH1 cells were HBV negative with slight AFP expression (0.6 μg/L). G banding results indicated that EHH1 cells were aneuploid cells, primarily hyperdiploid. Tumors were formed in all the 10 mice subcutaneously inoculated with EHH1 cells. The pathological types and differentiation degrees of the newly formed tumors were identical to their parental hepatocellular carcinoma. Conclusion: The established EHH1 cell line in this study maintaines the biologic characteristics of its original carcinoma, and might be a stable cell line of human hepatocellular carcinoma.
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Project Supported:
Supported by the National Natural Science Foundation of China (No.30728028)
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