Establishment and Biological Characterization of a Panel of Glioblastoma Multiforme (GBM) and GBM Variant Oncosphere Cell Lines
Authors: Brent A Orr
Amin B Kassam
Gary L Gallia
Charles G Eberhart
Jon D Weingart
Alessandro Olivi
Kelli M Wilson
Chetan Bettegowda
I-Mei Siu
Gregory J Riggins
Henry Brem
Vafi Salmasi
Alfredo Quinones-Hinojosa
Zev A Binder
Michael Lim
Journal: PloS one (2016)
Discipline: Multidisciplinary
Keywords: Tumor cell lines; Suspension cell lines; Glioblastoma multiforme; Neural stem cell culture; Genetic profiles
Affiliations: Department of Neurosurgery, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America., Johns Hopkins Physical Science Oncology Center and Institute for NanoBioTechnology, Johns Hopkins University, Baltimore, MD, United States of America., Department of Pathology, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America., Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, United States of America., Department of Neurosurgery, Aurora Neuroscience Innovation Institute, Milwaukee, WI, United States of America.

Abstract:  Objective: Human tumor cell lines form the basis of the majority of present day laboratory cancer research. These models are vital to studying the molecular biology of tumors and preclinical testing of new therapies. When compared to traditional adherent cell lines, suspension cell lines recapitulate the genetic profiles and histologic features of glioblastoma multiforme (GBM) with higher fidelity. Using a modified neural stem cell culture technique, here we report the characterization of GBM cell lines including GBM variants. Methods: Tumor tissue samples were obtained intra-operatively and cultured in neural stem cell conditions containing growth factors. Tumor lines were characterized in vitro using differentiation assays followed by immunostaining for lineage-specific markers. In vivo tumor formation was assayed by orthotopic injection in nude mice. Genetic uniqueness was confirmed via short tandem repeat (STR) DNA profiling. Results: Thirteen oncosphere lines derived from GBM and GBM variants, including a GBM with PNET features and a GBM with oligodendroglioma component, were established. All unique lines showed distinct genetic profiles by STR profiling. The lines assayed demonstrated a range of in vitro growth rates. Multipotency was confirmed using in vitro differentiation. Tumor formation demonstrated histologic features consistent with high grade gliomas, including invasion, necrosis, abnormal vascularization, and high mitotic rate. Xenografts derived from the GBM variants maintained histopathological features of the primary tumors. Conclusions: We have generated and characterized GBM suspension lines derived from patients with GBMs and GBM variants. These oncosphere cell lines will expand the resources available for preclinical study. 

Retracted: No

PubPeer comments:
 In the last 3 years I have tried 3 times to contact the authors to ask them 3 questions on the cell lines that they have established and that they describe in their very nice paper. As I never received an answer I leave these 3 questions in this PubPeer comment in the hope it may get answered! * 1) Can you provide the doubling time of these cell lines (they are not reported in the article but just described as slow or fast). * 2) Can you provide the age at sampling for the patients. * 3) For JHH-505 I annotated in the relevant Cellosaurus entries both  NCIt; C3058; Glioblastoma and  NCIt; C3288; Oligodendroglioma . Should I only put one of the two? If yes, which one? The relevant Cellosaurus entries are the following: https: web.expasy.org cgi-bin cellosaurus search?input=27028405 ,