Establishment of primary cell culture and an intracranial xenograft model of pediatric ependymoma: a prospect for therapy development and understanding of tumor biology
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Lorena Favaro Pavon1, Tatiana Tais Sibov1, Silvia Regina Caminada de Toledo2, Daniela Mara de Oliveira3, Francisco Romero Cabral4, Jean Gabriel de Souza5, Pamela Boufleur5, Luciana C. Marti4,6, Jackeline Moraes Malheiros7, Edgar Ferreira da Cruz8, Fernando F. Paiva9, Suzana M.F. Malheiros1,4, Manoel A. de Paiva Neto1, Alberto Tannús9, Sérgio Mascarenhas de Oliveira9, Nasjla Saba Silva2, Andrea Maria Cappellano2, Antonio Sérgio Petrilli2, Ana Marisa Chudzinski-Tavassi5 and Sérgio Cavalheiro1,2
1Department of Neurology and Neurosurgery, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil
2Pediatric Oncology Institute, Grupo de Apoio ao Adolescente e à Criança com Câncer (GRAACC), Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil
3Department of Genetics and Morphology, Universidade de Brasília, Brasília, Brazil
4Hospital Israelita Albert Einstein (HIAE), São Paulo, Brazil
5Biochemistry and Biophysics Laboratory, Butantan Institute, São Paulo, Brazil
6Allergy and Immunopathology Graduate Program, Faculdade de Medicina, Universidade de São Paulo (USP), São Paulo, Brazil
7Department of Physiology, Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil
8Discipline of Nephrology, Escola Paulista de Medicina (EPM), Universidade Federal de São Paulo (UNIFESP), São Paulo, Brazil
9São Carlos Institute of Physics, Universidade de São Paulo (USP), São Paulo, Brazil
Keywords: primary culture EPN cells; pluripotency markers; animal model; MRI; preclinical studies
Received: November 15, 2017 Accepted: March 06, 2018 Published: April 24, 2018
Background: Ependymoma (EPN), the third most common pediatric brain tumor, is a central nervous system (CNS) malignancy originating from the walls of the ventricular system. Surgical resection followed by radiation therapy has been the primary treatment for most pediatric intracranial EPNs. Despite numerous studies into the prognostic value of histological classification, the extent of surgical resection and adjuvant radiotherapy, there have been relatively few studies into the molecular and cellular biology of EPNs.
Results: We elucidated the ultrastructure of the cultured EPN cells and characterized their profile of immunophenotypic pluripotency markers (CD133, CD90, SSEA-3, CXCR4). We established an experimental EPN model by the intracerebroventricular infusion of EPN cells labeled with multimodal iron oxide nanoparticles (MION), thereby generating a tumor and providing a clinically relevant animal model. MRI analysis was shown to be a valuable tool when combined with effective MION labeling techniques to accompany EPN growth.
Conclusions: We demonstrated that GFAP/CD133+CD90+/CD44+ EPN cells maintained key histopathological and growth characteristics of the original patient tumor. The characterization of EPN cells and the experimental model could facilitate biological studies and preclinical drug screening for pediatric EPNs.
Methods: In this work, we established notoriously challenging primary cell culture of anaplastic EPNs (WHO grade III) localized in the posterior fossa (PF), using EPNs obtained from 1 to 10-year-old patients (n = 07), and then characterized their immunophenotype and ultrastructure to finally develop a xenograft model.
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