RESUMEN
Brain tumours are the leading cause of cancer mortality in children and remain difficult to cure despite advances in surgery and adjuvant therapy. To identify cancer stem-like cells in established cell lines from three paediatric brain tumour [PBT] gliomas. Queen's Medical Centre/ Nottingham city/ UK. Three glioma cell lines were studied including one high grade glioblastoma multiforme [BT4], one well differentiated oligodendroglioma [Olig1], and one recurrent ependymoma [EPN1]. A control cell line of mouse neural stem cells [C17.2] was also included for comparison. Established tumour cell lines maintain stem cell marker [nestin and Sox2] expression when grown as monolayers in 15% foetal bovine serum/Dulbecco's modified Eagle's medium. Cells derived from these cell lines are able to form neurospheres when cultured in serum-free stem cell media containing basic fibroblast growth factor and human epidermal growth factor. These neurospheres are self-renewable and re-form new neurospheres when dissociated and cultured in fresh medium supplemented with growth factors. The percentages of neural stem cell marker CD 133 positive cells were determined by flow cytometry analysis of neurospheres from the three cultured cell lines, BT4 47.2% +/- 10.5, EPN1 40.4% +/- 8.9 and Olig1 48.7% +/- 2.9 [mean +/- standard error]. Under conditions promoting differentiation, cells derived from neurospheres were multipotent giving rise to neurons, astrocytes, and oligodendrocytes, at different levels for each tumour cell line. Paediatric gliomas contain cancer stem-like cells that are able to self-renew and differentiate into three neural lineages