Human recombinant bfgf and egf

Parts of the surgical samples were enzymatically dissociated into single cells, following the procedures previously reported [13] (link). Dissociated GBM cells were cultured in neurobasal media with N2 and B27 supplements (0.5 each; Invitrogen) and human recombinant bFGF and EGF (25 ng/ml each; R&D Systems) (NBE condition).

Animal experiments were approved by the Institutional Review Board and conducted in accordance with the “National Institutes of Health Guide for the Care and Use of Laboratory Animals” (NIH publication). Tumors were classified as GBM based on WHO criteria after review by pathologists. Parts of the surgical samples were enzymatically dissociated into single cells, following the procedures previously reported^{23 (link)}. Dissociated GBM cells were cultured in neurobasal media with N2 and B27 supplements (0.5X each; Invitrogen) and human recombinant bFGF and EGF (25 ng/ml each; R&D Systems) (NBE condition). Acutely dissociated GBM cells were stereotactically (2 mm left and 1 mm anterior to the bregma, 2 mm deep from the dura) injected into the brains of immune-compromised NOD/SCID Il2rg^−/− (NOG) mice within 12 hours after surgery (2.5 × 10⁴–1.0 × 10⁵ cells in 10 ml HBSS for each mice, n = 4–19 for each sample). Mice with the reduction of the total body weight (>20%) were sacrificed, and brains were processed for paraffin or frozen section.