Isolation and functional characterization of primary endothelial cells from mouse cerebral cortex

Ouellette, Julie; Lacoste, Baptiste

doi:10.1016/j.xpro.2021.101019

Cited by 2 publications

(2 citation statements)

References 5 publications

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“…VIII in the Supplement). While Flk1 expression was observed in Gp38 + cells, we did not observe CD31 expression, indicating that conditions used in our culture conditions supports cell expansion, but are not optimized for full endothelial differentiation consistent with previous studies that have shown that endothelial differentiation requires specific cytokines as well as high density and specific substrate conditions 74 , 75 , which we could not used in tandem with our colony formation assays. We further confirmed the ability to form colonies at the single cell level (Fig.…”

Section: Resultssupporting

confidence: 86%

Hypoxia promotes a perinatal-like progenitor state in the adult murine epicardium

Sayed

Turoczi

Soares-da-Silva

et al. 2022

Sci Rep

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The epicardium is a reservoir of progenitors that give rise to coronary vasculature and stroma during development and mediates cardiac vascular repair. However, its role as a source of progenitors in the adult mammalian heart remains unclear due to lack of clear lineage markers and single-cell culture systems to elucidate epicardial progeny cell fate. We found that in vivo exposure of mice to physiological hypoxia induced adult epicardial cells to re-enter the cell cycle and to express a subset of developmental genes. Multiplex single cell transcriptional profiling revealed a lineage relationship between epicardial cells and smooth muscle, stromal cells, as well as cells with an endothelial-like fate. We found that physiological hypoxia promoted a perinatal-like progenitor state in the adult murine epicardium. In vitro clonal analyses of purified epicardial cells showed that cell growth and subsequent differentiation is dependent upon hypoxia, and that resident epicardial cells retain progenitor identity in the adult mammalian heart with self-renewal and multilineage differentiation potential. These results point to a source of progenitor cells in the adult heart that can be stimulated in vivo and provide an in vitro model for further studies.

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Section: Resultssupporting

confidence: 86%

Hypoxia promotes a perinatal-like progenitor state in the adult murine epicardium

Sayed

Turoczi

Soares-da-Silva

et al. 2022

Sci Rep

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“…The embedded cortex is then sectioned tangentially with a vibratome (step 2), stained, and mounted (see also Refs. 5 and 12 ).…”

Section: Methodsmentioning

confidence: 98%

Unbiased analysis of mouse brain endothelial networks from two- or three-dimensional fluorescence images

et al. 2022

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. Significance: A growing body of research supports the significant role of cerebrovascular abnormalities in neurological disorders. As these insights develop, standardized tools for unbiased and high-throughput quantification of cerebrovascular structure are needed. Aim: We provide a detailed protocol for performing immunofluorescent labeling of mouse brain vessels, using thin ( ) or thick (50 to ) tissue sections, followed respectively by two- or three-dimensional (2D or 3D) unbiased quantification of vessel density, branching, and tortuosity using digital image processing algorithms. Approach: Mouse brain sections were immunofluorescently labeled using a highly selective antibody raised against mouse Cluster of Differentiation-31 (CD31), and 2D or 3D microscopy images of the mouse brain vasculature were obtained using optical sectioning. An open-source toolbox, called Pyvane, was developed for analyzing the imaged vascular networks. The toolbox can be used to identify the vasculature, generate the medial axes of blood vessels, represent the vascular network as a graph, and calculate relevant measurements regarding vascular morphology. Results: Using Pyvane, vascular parameters such as endothelial network density, number of branching points, and tortuosity are quantified from 2D and 3D immunofluorescence micrographs. Conclusions: The steps described in this protocol are simple to follow and allow for reproducible and unbiased analysis of mouse brain vascular structure. Such a procedure can be applied to the broader field of vascular biology.

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Isolation and functional characterization of primary endothelial cells from mouse cerebral cortex

Cited by 2 publications

References 5 publications

Hypoxia promotes a perinatal-like progenitor state in the adult murine epicardium

Hypoxia promotes a perinatal-like progenitor state in the adult murine epicardium

Unbiased analysis of mouse brain endothelial networks from two- or three-dimensional fluorescence images

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