2.2. Patient-Derived GBM Cells
GBM 3691 cells were obtained as de-identified specimens that were
initially acquired as primary human brain tumor patient specimens in
accordance with appropriate, approved Institutional Review Board (IRB)
protocols. GBM 3691 was a kind gift from Dr. Jeremy Rich (University of
Pittsburgh; IRBs from Duke University and The Cleveland Clinic;
transferred via MTAs). Cells were cultured in suspension at 37°C at 5%
CO2 in Neurobasal media (minus phenol red; Gibco) with
added B27 (minus Vitamin A; Gibco), human fibroblast growth factor-2 (10
ng/mL; Miltenyi Biotec), human epidermal growth factor (10 ng/mL;
Miltenyi Biotec), L-glutamine (2 mM; Gibco), sodium pyruvate (1 mM;
Gibco), and penicillin/streptomycin (100 I.U./ml/100 ug/ml; Gibco).
Prior to seeding within hydrogels, cell aggregates are dissociated into
a single cell suspension using TrypLE (Gibco).