A total of six pairs of neurospheres derived from both GBM and peritumoral tissue (at a distance < 1 cm from macroscopic tumor border), called Glioblastoma Cancer Stem Cells (GCSCs) and Peritumoral Cancer Stem Cells (PCSCs), respectively, were kindly provided by Vescovi and Binda. According to the classification of human gliomas by WHO [29 (link)], we analyzed the GCSC and PCSC neurospheres for the mutational status of IDH1 [30 (link), 31 (link)]. Mutational IDH1/2 status and MGMT promoter methylation status were also investigated in all the available tissue samples from which the neurospheres were derived [32 (link)].
The neurospheres were cultured in NeuroCult™ NS-A Proliferation Kit (StemCell Technologies Inc, Vancouver, BC, Canada) supplemented with 20 ng/mL human recombinant EGF, 10 ng/ml human recombinant bFGF and 2 µg/mL heparin (all from StemCell Technologies Inc.), as described previously [33 (link)]. All cell cultures were maintained at 37 °C in a 5% CO2 humidified atmosphere.
The neurospheres were cultured in NeuroCult™ NS-A Proliferation Kit (StemCell Technologies Inc, Vancouver, BC, Canada) supplemented with 20 ng/mL human recombinant EGF, 10 ng/ml human recombinant bFGF and 2 µg/mL heparin (all from StemCell Technologies Inc.), as described previously [33 (link)]. All cell cultures were maintained at 37 °C in a 5% CO2 humidified atmosphere.
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