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Isolation, cultivation and identification of cancer stemlike cells from human liver cancer tissues
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Abstract:
Objective:To isolate and cultivate liver cancer stemlike cells (hLCSLCs) from human liver cancer specimens, so as to lay a foundation for stem celltargeted therapy of liver cancer. Methods: The hLCSLCs were obtained from fresh liver cancer tissue using enzymatic digestion and shortterm primary culture. Serumfree medium suitable for suspension sphere formation of hLCSLCs was selected by culturing them with serumfree medium supplement with heparin, albumin or hydrocortisone. Numbers of side population (SP) cells, and expressions of CD133 and CD90 in the hLCSLCs and sphere cells were examined by flow cytometry assay. Tumor formation ability of hLCSLCs and sphere cells were assessed by tumor formation assay. Results: hLCSLCs were successfully obtained from human liver cancer tissues, with SP cells being 0.9%, CD133 positive cells being 0.8% and CD90 positive cells being 12.7% in the hLCSLCs. The tumor formation rate was 100% when 2 000 SP cells were subcutaneously injected into nude mice. Serumfree medium containing heparin was more suitable for sphere formation of hLCSLCs, with SP cells being 4.6%, CD133+ cells being 9.7%, and CD90+ cells being 48% in these sphere cells. The tumor formation rate was 100% when 10 000 sphere cells were injected into nude mice. Conclusion: We have successfully established a method for isolating hLCSLCs from human liver cancer tissues. Serumfree suspension medium containing heparin can effectively enrich hLCSLCs .
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Project Supported:
Supported by the Major Basic Research and Development Program(973) of China（No.2009CB521804）
Clc Number:
