Establishment of Human chordoma cell line HBC2

Objective To establish human chordoma cell lines.Methods The tumor tissue samples were obtained from surgery.The tumor cells were mechanically dissociated and purified based on the different rates of attachment among various cell types.The cells were cultured in DMEM/F-12 medium and passaged in vitro.The morphology and ultrastructure of tumor cells (passage 15) were observed by microscope and electron microscope.The proteins S100,Galectin-3,Fascin-1and Brachyury were measured by histochemical staining.Four phases of the cell cycle was analyzed by the flow cytometer.The karyotype analysis of tumor cells was performed.The tumor cells were subcutaneously injected into the nude mice.Results The HBC2 cell line was characterized by prominent vacuoles of mucus pushing the nuclei to the side.The proteins S100,Galectin-3,Fascin-1 and Brachyury were positively stained in the cell line.The flow cytometer showed 51.3％ of the cells were at G1 phase,23.6％ at S,and 25.0％ at G2-M.The value of G2/G1 was almost 2.The heteroploid karyotype of the cells was indicated.The tumor formation in nude mice was found by HBC2 cells thansplantation.Conclusions Human chordoma cell line HBC2 was successfully established and cultured in vitro on the basis of maintaining its characteristics of chordoma in the process of passage. Key words: Chordoma;  Skull base ;  Cell line ;  Karyotype analysis ;  Umorigenicity