Characterization of Human Glioblastoma Cell Lines

The human glioblastoma cell lines LN229 (ATCC-CRL-2611), U251 (ECACC 89081403; formerly known as U373MG), U87MG (ECACC 89081402), and T98G (ECACC No. 92090213) were obtained from the European Collection of Authenticated Cell Cultures (ECACC, Salisbury, UK) or the American Type Culture Collection (ATCC, Manassas, VA, USA) and cultured as described previously [24 (link)]. Human primary GBM cultures (n = 2) were produced by dissociation and cultured according to established techniques as described before [24 (link)]. Human primary GBM stem-like cell cultures (n = 8) as well as GBM cell line-derived stem-like cells were established and intensively characterized by the formation of neurospheres, the ability to survive and proliferate under stem cell conditions, and the ability to differentiate into more mature cells as described before [3 (link),25 (link),26 (link),27 (link)]. The purity of the GBM cells was ascertained by immunostaining with cell type-specific markers and by the absence of contamination with mycoplasms. GBM cell line identity was verified by short tandem repeat profiling at the Department of Forensic Medicine (Kiel, Germany) using the Powerplex HS Genotyping Kit (Promega, Madison, WI, USA) and the 3500 Genetic Analyzer (Thermo Fisher Scientific, Waltham, MA, USA) as previously described [3 (link)].