Derivation and Culture of Human Glioblastoma Neurosphere Lines

The human glioblastoma neurosphere lines HSR-GBM1A (20913) and HSR-GBM 1B (10627) were originally derived by Vescovi and colleagues and maintained in serum-free medium supplemented with epidermal growth factor and fibroblast growth factor (Vescovi et al., 1999 (link); Galli et al., 2004 (link); Bar et al., 2007 (link); Sun et al., 2009 (link)). Cells were incubated in a humidified incubator containing 5% CO₂/95% air at 37 °C, and passaged every 4–5 days. The GBM-DM140207 and GBM-KK190156 neurosphere lines were derived from glioblastomas at the University of Freiburg and kindly provided by Dr Jaroslaw Maciaczy. The primary neurospheres JHH551 was derived from a malignant glioma at Johns Hopkins using the same methods and culture conditions as described in Galli et al. (2004) (link). JHH551 neurospheres were used at passage ≤10. All human materials were obtained and used in compliance with the Johns Hopkins IRB.