An adapted dorsal skinfold model used for 4D intravital followed by whole-mount imaging to reveal endothelial cell–pericyte association

Ann L.B. Seynhaeve; Timo L.M. ten Hagen

doi:10.1038/s41598-021-99939-w

An adapted dorsal skinfold model used for 4D intravital followed by whole-mount imaging to reveal endothelial cell–pericyte association

Ann L.B. Seynhaeve^*, Timo L.M. ten Hagen

^*Corresponding author for this work

Pathology

Research output: Contribution to journal › Article › Academic › peer-review

11 Citations (Scopus)

Abstract

Endothelial cells and pericytes are highly dynamic vascular cells and several subtypes, based on their spatiotemporal dynamics or molecular expression, are believed to exist. The interaction between endothelial cells and pericytes is of importance in many aspects ranging from basic development to diseases like cancer. Identification of spatiotemporal dynamics is particularly interesting and methods to studies these are in demand. Here we describe the technical details of a method combining the benefits of high resolution intravital imaging and whole-mount histology. With intravital imaging using an adapted light weight dorsal skinfold chamber we identified blood flow patterns and spatiotemporal subtypes of endothelial cells and pericytes in a 4D (XYZ, spatial+T, time dimension) manner as representative examples for this model. Thereafter the tissue was extracted and stained as a whole-mount, by which the position and volumetric space of endothelial cells as well as pericytes were maintained, to identify molecular subtypes. Integration of the two imaging methods enabled 4D dissection of endothelial cell–pericyte association at the molecular level.

Original language	English
Article number	20389
Journal	Scientific Reports
Volume	11
Issue number	1
Early online date	14 Oct 2021
DOIs	https://doi.org/10.1038/s41598-021-99939-w
Publication status	Published - Dec 2021

Bibliographical note

Funding Information:
We like to thank Rien van Haperen for the creation of the eNOStag-GFP animal line, Hanna Eilken, Susanne Adams and Ralf Adams for the PdgfrbCreERT2 line and the animal care takers for their help. The microscopy facilities used are part of the Erasmus Optical Imaging Center and we like to thank Gert van Cappellen, Gert-Jan Kremers and the other staff members of the eOIC for their service. We also thank Alex Brouwer and his colleagues of the EMI for the development and manufacturing of the window materials and the adaptation of the microscope and prof. dr. Stallcup for donating his antibodies. This study was supported by the Dutch Cancer Society, “Vereniging Trustfonds” Erasmus University Rotterdam, and “Stichting Erasmus Heelkundig Kankeronderzoek”.

Publisher Copyright:
© 2021, The Author(s).

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Cite this

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abstract = "Endothelial cells and pericytes are highly dynamic vascular cells and several subtypes, based on their spatiotemporal dynamics or molecular expression, are believed to exist. The interaction between endothelial cells and pericytes is of importance in many aspects ranging from basic development to diseases like cancer. Identification of spatiotemporal dynamics is particularly interesting and methods to studies these are in demand. Here we describe the technical details of a method combining the benefits of high resolution intravital imaging and whole-mount histology. With intravital imaging using an adapted light weight dorsal skinfold chamber we identified blood flow patterns and spatiotemporal subtypes of endothelial cells and pericytes in a 4D (XYZ, spatial+T, time dimension) manner as representative examples for this model. Thereafter the tissue was extracted and stained as a whole-mount, by which the position and volumetric space of endothelial cells as well as pericytes were maintained, to identify molecular subtypes. Integration of the two imaging methods enabled 4D dissection of endothelial cell–pericyte association at the molecular level.",

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An adapted dorsal skinfold model used for 4D intravital followed by whole-mount imaging to reveal endothelial cell–pericyte association

Abstract

Bibliographical note

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