Establishment and Characterization of Primary Glioblastoma Cell Lines from Fresh and Frozen Material: A Detailed Comparison

<div><p>Background</p><p>Development of clinically relevant tumor model systems for glioblastoma multiforme (GBM) is important for advancement of basic and translational biology. High molecular heterogeneity of GBM tumors is well recognized, forming the rationale for molecular tests required before administration of several of the novel therapeutics rapidly entering the clinics. One model that has gained wide acceptance is the primary cell culture model. The laborious and time consuming process is rewarded with a relative high success rate (about 60%). We here describe and evaluate a very simple cryopreservation procedure for GBM tissue prior to model establishment that will considerably reduce the logistic complexity.</p><p>Methods</p><p>Twenty-seven GBM samples collected ad hoc were prepared for primary cell culture freshly from surgery (#1) and after cryopreservation (#2).</p><p>Results</p><p>Take rates after cryopreservation (59%) were as satisfactory as from fresh tissue (63%; p = 1.000). We did not observe any relevant molecular or phenotypic differences between cell lines established from fresh or vitally frozen tissue. Further, sensitivity both towards standard chemotherapeutic agents (Temozolomide, BCNU and Vincristine) and novel agents like the receptor tyrosine kinase inhibitor Imatinib did not differ.</p><p>Conclusions</p><p>Our simple cryopreservation procedure facilitates collection, long-time storage and propagation (modeling) of clinical GBM specimens (potentially also from distant centers) for basic research, (pre-) clinical studies of novel therapies and individual response prediction.</p></div

IC₅₀ values.

<p>Calculated IC₅₀ values for all cell line pairs and substances tested are listed.</p

Primers used for mutation analyses.

<p>Primers used for mutation analyses.</p

Doubling times.

<p>Mean doubling times for the cell lines are displayed placing the pairs side by side. The doubling time for each cell line was determined three times; the mean doubling time (in h) and standard deviation were calculated and depicted in a bar chart.</p

Clinical information.

<p>Relevant clinical patient data concerning age (at time point of surgery), gender (M = male; F = female), tumor localization, further information (if provided) and survival in months († = patient died; bold = patient still alive on January 25^th 2013) are summarized.</p

Cell line morphology.

<p>Phenotypes of the cell lines captured by microphotography are displayed pairwise.</p