A versatile ex vivo technique for assaying tumor angiogenesis and microglia in the brain
Autor: | Tina Sehm, Ilker Y. Eyüpoglu, Ali Ghoochani, Zheng Fan, Stefan W. Hock, Michael Buchfelder, Nicolai E. Savaskan, Eduard Yakubov |
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Jazyk: | angličtina |
Rok vydání: | 2015 |
Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Angiogenesis Green Fluorescent Proteins Brain tumor slice culture Mice Transgenic Biology 03 medical and health sciences angiogenesis Slice preparation Organ Culture Techniques In vivo Glioma Cell Line Tumor medicine Temozolomide Tumor Microenvironment Animals Humans Rats Wistar Antineoplastic Agents Alkylating Cell Proliferation Tumor microenvironment Microscopy Confocal Microglia Neovascularization Pathologic Brain Neoplasms neuronal cell death glioblastoma Reproducibility of Results medicine.disease Cell biology Dacarbazine 030104 developmental biology medicine.anatomical_structure Oncology Microscopy Fluorescence ex vivo Ex vivo Research Paper |
Zdroj: | Oncotarget |
ISSN: | 1949-2553 |
Popis: | Primary brain tumors are hallmarked for their destructive activity on the microenvironment and vasculature. However, solely few experimental techniques exist to access the tumor microenvironment under anatomical intact conditions with remaining cellular and extracellular composition. Here, we detail an ex vivo vascular glioma impact method (VOGIM) to investigate the influence of gliomas and chemotherapeutics on the tumor microenvironment and angiogenesis under conditions that closely resemble the in vivo situation. We generated organotypic brain slice cultures from rats and transgenic mice and implanted glioma cells expressing fluorescent reporter proteins. In the VOGIM, tumor-induced vessels presented the whole range of vascular pathologies and tumor zones as found in human primary brain tumor specimens. In contrast, non-transformed cells such as primary astrocytes do not alter the vessel architecture. Vascular characteristics with vessel branching, junctions and vessel meshes are quantitatively assessable as well as the peritumoral zone. In particular, the VOGIM resembles the brain tumor microenvironment with alterations of neurons, microglia and cell survival. Hence, this method allows live cell monitoring of virtually any fluorescence-reporter expressing cell. We further analyzed the vasculature and microglia under the influence of tumor cells and chemotherapeutics such as Temozolamide (Temodal/Temcad®). Noteworthy, temozolomide normalized vasculare junctions and branches as well as microglial distribution in tumor-implanted brains. Moreover, VOGIM can be facilitated for implementing the 3Rs in experimentations. In summary, the VOGIM represents a versatile and robust technique which allows the assessment of the brain tumor microenvironment with parameters such as angiogenesis, neuronal cell death and microglial activity at the morphological and quantitative level. |
Databáze: | OpenAIRE |
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