The study of cell proliferation and glioma spheroid formation from human glioma cell line U87

• Main Authors: Chen Chieh Liao, 廖振捷
• Other Authors: Chun-Che Chen
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/22253292072873864148

碩士 === 國立嘉義大學 === 水生生物科學系研究所 === 97 === Glioma is one of the most common primary brain tumors in human. Like other human malignancies, several glioma cells consist with complex genetic backgrounds. There are neural stem cell (NSC)-like cells or tumor stem cells in the glioma lesion site. NSCs are characterized as being self-renewing, immature, undifferentiated, and multipotent, meaning that they can differentiate as neurons, astrocytes, and oligodendrocytes. The human glioblastoma cell line U87 is one of the widely used cell lines in the studies of brain tumors. In this study, we found that U87 cell which with several characteristics that could form glioma spheroids and the capability of self-renewal in vitro. We observed that cell can aggregate and form glioma spheroid from monolayer. In order to examine the effect of cell density on cell proliferation and glioma spheroid development. Cells were seeded in medium with different cell densities: 1x105 , 2x105 , 5x105 , 8x105 cells/ml. The forming glioma spheroids is more efficient in higher density seeding of U87 cells during 6 days of observation period. Glioma spheroid developed that grew through cell proliferation and reached diameters from 200 to 720 μm. The diameters of glioma spheroids are the most from 300 to 450 μm. Time-lapses microscopy is used for the time series observation for cell aggregation and self-renewal from single cell to spheroid and also migrated from spheroid to monolayer. The results show U87 cell can aggregate start from the 4th hour after seeding , and become glioma spheroids at the 44th hour. The new cell migrated at the forth hours after seeding single glioma spheroid. These cells possessed the ability of self-renewal and multipotency , which can be defined as the characteristics of CSCs. The proteomic of U87 cell, we found some proteins have high expression level in glioma spheroid. These proteins such as keratin 9/10, vimentin and pericentrin. When forming glioma spheroid the structure change case by these proteins.