Organotypic 3D Ex Vivo Co-culture Model of the Macro-metastasis/Organ Parenchyma Interface.
Methods Mol Biol
; 2764: 165-176, 2024.
Article
em En
| MEDLINE
| ID: mdl-38393595
ABSTRACT
The macro-metastasis/organ parenchyma interface (MMPI) is gaining increasing significance due to its prognostic relevance for cancer (brain) metastasis. We have developed an organotypic 3D ex vivo co-culture model that mimics the MMPI and allows us to evaluate the histopathological growth pattern (HGP) and infiltration grade of the tumor cells into the neighboring brain tissue and to study the interactions of cancer and glial cells ex vivo. This system consists of a murine brain slice and a 3D tumor plug that can be co-cultured for several days. After slicing the brain of 5- to 8-day-old mice, a Matrigel plug containing fluorescent-labelled tumor cells is placed next to it, so that tumor cells in the 3D plug and glial cells in the brain slice can interact at the interface for up to 96 h. To facilitate the positioning of the co-culture and increase the reproducibility of the model, a brain spacer can be used. The HGP and infiltration of the tumor cells into the brain slice as well as the activation of the glial cells can be assessed by live and/or confocal microscopy after immunofluorescence staining of microglia and/or astrocytes. Alternatively, the co-culture can also be used for other purposes, such as RNA analysis. This organotypic 3D ex vivo co-culture offers a perfect tool for preliminary screenings before in vivo experiments and reduces the number of animals, thus contributing to the 3R concept as a central precept in preclinical research.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Neoplasias Encefálicas
/
Neuroglia
Limite:
Animals
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Alemanha