Human gliomas harbour cancer stem cells (CSCs) that evolve along the course of the disease, forming highly heterogeneous subpopulations within the tumour mass. These cells possess self-renewal properties and appear to contribute to tumour initiation, metastasis and resistance to therapy. CSC cultures isolated from surgical samples are considered the best preclinical in vitro model for primary human gliomas. However, it is not yet well characterized to which extent their biological and functional properties change during in vitro passaging in the serum-free culture conditions. Here, we demonstrate that our CSC-enriched cultures harboured from one to several CSC clones from the human glioma sample. When xenotransplanted into mouse brain, these cells generated tumours that reproduced at least three different dissemination patterns found in original tumours. Along the passages in culture, CSCs displayed increased expression of stem cell markers, different ratios of chromosomal instability events, and a varied response to drug treatment. Our findings highlight the need for better characterization of CSC-enriched cultures in the context of their evolution in vitro, in order to uncover their full potential as preclinical models in the studies aimed at identifying molecular biomarkers and developing new therapeutic approaches of human gliomas.

Centro Nacional de Biotecnología Madrid Spain

Division of Biopharmaceutics and Pharmacokinetics University of Helsinki Helsinki Finland

Fundación de Investigación HM Hospitales HM Hospitales Madrid Spain

Genetic and Genomic Unit Fundación Centro de Investigación Príncipe Felipe Valencia Spain

Instituto de Investigaciones Biomédicas CIBERER CSIC UAM Madrid Spain

Instituto de Medicina Molecular Aplicada School of Medicine San Pablo CEU University Campus de Montepríncipe Madrid Spain

Instituto Madrileño de Estudios Avanzados IMDEA Nanociencia Madrid Spain

International Clinical Research Center Center for Translational Medicine St Anne's University Hospital Brno Czech Republic

Neuro Oncology Unit Instituto de Salud Carlos 3 UFIEC Madrid Spain

Neurosurgery Department Hospital Universitario la Fe de Valencia Valencia Spain

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