Density gradient centrifugation cells and metastatic liver cancer cell lines established.docVIP

 PAGE \* MERGEFORMAT 22 Density gradient centrifugation cells and metastatic liver cancer cell lines established Authors: Zhang Meng, Hu Shao-wei, ginger orange, Wen Jian-ming 【Abstract】 【Objective】 establishment of a new hepatocellular carcinoma cell lines, in order to provide more experimental study purified liver cancer. 【Methods】 from the primary tumor in patients with hepatocellular carcinoma and portal vein tumor thrombus to obtain cancer tissue, using Percoll density gradient separation were cultured cancer cells, using micro-environment for cell digestion method and the establishment of cloned cell lines, cultured cells, the success of growth curve of the Department of the determination of G-banded karyotypes of chromosome analysis, comparative genomic hybridization, and nude mice inoculated into the tumor experiments. 【Results】 The 20 cases of liver cancer cell isolation and culture organizations, and cloning, two cases of successfully established cell lines H2M and H4M, these two cells are from portal vein tumor thrombus, and their chromosome numbers were measured 3 times super-body (71 ~ 78). CGH tests showed that the major cytogenetic H2M changed to chromosome 4q, 13q, 16q, 17p, and 19p deletion and 1q, 3q, 5p, 6p, 7q, and 8q amplification, which contains a marker chromosome 1q, and 6p [t (1 ; 6) chromosome]; while H4M chromosome 8p, 9, 13q, 16q deletion and 6p, 7p, 11p, 11q13 amplification, including a marker chromosome containing a long homogeneously staining area (hsr). H2M cells in nude mice inoculated 1 month, resulting in a typical human hepatocellular carcinoma, but not yet into the tumor H4M inoculation. 【Conclusion】 Percoll density gradient separation of cancer cells and the microenvironment of cancer cells to obtain clone digestion method is to establish primary cell lines an effective manner. 2 cases of liver cancer cell lines for the future liver cancer study has provided experimental material, the measured liver cancer cell