Stem cell cultures derived from pediatric brain tumors accurately model the originating tumors.
Brain tumors are the leading cause of cancer-related death in children but high-grade gliomas in children and adolescents have remained a relatively under-investigated disease despite this. A better understanding of the cellular and molecular pathogenesis of the diseases is required in order to improve the outcome for these children. In vitro-cultured primary tumor cells from patients are indispensable tools for this purpose by enabling functional analyses and development of new therapies. However, relevant well-characterized in vitro cultures from pediatric gliomas cultured under serum-free conditions have been lacking. We have therefore established patient-derived in vitro cultures and performed thorough characterization of the cells using large-scale analyses of DNA methylation, copy-number alterations and investigated their stability during prolonged time in culture. We show that the cells were stable during prolonged culture in serum-free stem cell media without apparent alterations in morphology or growth rate. The cells were proliferative, positive for stem cell markers, able to respond to differentiation cues and initiated tumors in zebrafish and mice suggesting that the cells are cancer stem cells or progenitor cells. The cells accurately mirrored the tumor they were derived from in terms of methylation pattern, copy number alterations and DNA mutations. These unique primary in vitro cultures can thus be used as a relevant and robust model system for functional studies on pediatric brain tumors.
Department of Pathology, Sahlgrenska Cancer Center, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Anna
A
Wenger
Department of Pathology, Sahlgrenska Cancer Center, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Susanna
S
Larsson
Department of Oncology, Sahlgrenska Cancer Center, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Anna
A
Danielsson
Department of Pathology, Sahlgrenska Cancer Center, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Kirstine Juul
KJ
Elbæk
Department of Psychiatry and Neurochemistry, Institute of Neuroscience and Physiology, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.|Department of Neuropathology, Nuffield Department of Clinical Neurosciences, University of Oxford, John Radcliffe Hospital, Oxford, United Kingdom.
Petronella
P
Kettunen
Department of Clinical Neuroscience and Rehabilitation, Institute of Neuroscience and Physiology, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Magnus
M
Tisell
Department of Pediatrics, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.|The Queen Silvia Children's Hospital, Sahlgrenska University Hospital, Gothenburg, Sweden.
Magnus
M
Sabel
Department of Pediatrics, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.|The Queen Silvia Children's Hospital, Sahlgrenska University Hospital, Gothenburg, Sweden.
Birgitta
B
Lannering
Department of Pathology, Sahlgrenska University Hospital, Gothenburg, Sweden.
Claes
C
Nordborg
Department of Pathology, Sahlgrenska Cancer Center, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.|The Queen Silvia Children's Hospital, Sahlgrenska University Hospital, Gothenburg, Sweden.
Elizabeth
E
Schepke
Department of Pathology, Sahlgrenska Cancer Center, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
Helena
H
Carén
D000818:Animals; D001932:Brain Neoplasms / Q000235:genetics / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods*; D002648:Child; D020732:Cytogenetic Analysis; D005434:Flow Cytometry; D064593:Heterografts; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D051379:Mice; D014411:Neoplastic Stem Cells / Q000473:pathology*; D015027:Zebrafish
DNA methylation; cancer stem cells; glioblastoma; immunodeficient mice; pediatric