Surprisingly in vitro conditions allowing to culture glioblastoma cells presenting EGFR amplification were not known until now. Our evaluation of EGFR amplification status in glioblastoma (GBM) culture demonstrated that this anomaly was preserved for months in spheroids (aggregated glioblastoma cells) at a level comparable to the earliest passage of cell culture. In contrast, and in accordance with already published data we detected it as completely lost in the adherent culture. Apparently glioblastoma cells presenting EGFR amplification become apoptotic in the regular cell culture conditions. In addition discrepant expression of SOX2 and multilineage phenotype recognized as a markers of neural progenitors was observed in monolayer and 3D culture. Moreover, our analyses showed a decreased invasion potential of adherent GBM loosing EGFR amplification, and spheroids maintaining EGFR amplification. In conclusion, our findings confirm that GBM-derived spheroids seem to be a promising tool to preserve original molecular features of the tumor in vitro, with a special emphasis on EGFR gene aberrations, including EGFRvIII, regarded as novel therapeutic target. Our last unpublished and preliminary data suggest that mechanism responsible for in vitro death of adherent glioblastoma cells showing EGFR amplification seems to be linked to the artificial in vitro cell-cell interaction rather than to the lack of proper autocrine effects. Those suggestion came from analysis of glioblastoma cells in artificial brain tissues system and analysis of aggregated glioblastoma cells only. The present study was supported by the Ministry of Science and Higher Education, Poland, grant No. N N401047337 and Foundation for Polish Science(PARENT-BRIDGE Programme - support for women, No.POMOST_C/15).
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