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Time-Lapse Observation of Cell Dynamics During Angiogenesis Using the Rat Mesentery Culture Model.

Authors
  • Lampejo, Arinola O1
  • Hodges, Nicholas A1
  • Rozenblum, Maximillian1
  • Murfee, Walter L2
  • 1 J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL, USA.
  • 2 J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, Gainesville, FL, USA. [email protected].
Type
Published Article
Journal
Methods in molecular biology
Publisher
Clifton, N.J. : Humana Press
Publication Date
Jan 01, 2024
Volume
2711
Pages
63–75
Identifiers
DOI: 10.1007/978-1-0716-3429-5_6
PMID: 37776449
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

The ability to track cells and their interactions with other cells during physiological processes offers a powerful tool for scientific discovery. An ex vivo model that enables real-time investigation of cell migration during angiogenesis in adult microvascular networks would enable observation of endothelial cell dynamics during capillary sprouting. Angiogenesis is defined as the growth of new blood vessels from existing ones and involves multiple cell types including endothelial cells, pericytes, and interstitial cells. The incorporation of these cell types in a physiologically relevant environment, however, represents a challenge for biomimetic model development. Recently, our laboratory has developed the rat mesentery culture model, which enables investigation of angiogenesis in an intact tissue. The objective of this chapter is to detail a protocol for tracking cellular dynamics during angiogenesis using the rat mesentery tissue culture model. The method involves harvesting mesentery tissues from adult SD-EGFP rats, culturing them in MEM + 10% fetal bovine serum, and imaging network regions over the time course of angiogenesis. In example applications, time-lapse comparison of microvascular networks in cultured tissues confirmed dramatic increases in GFP-positive capillary sprouting and GFP-positive segment density. Additionally, tracking of individual capillary sprout extensions revealed their ability to "jump" by disconnecting from one vessel segment and reconnecting to another segment in the network. GFP-positive sprouts were also capable of undergoing subsequent regression. The representative results support the use of the rat mesentery culture model for identifying and tracking cellular dynamics during angiogenesis in intact microvascular networks. © 2024. Springer Science+Business Media, LLC, part of Springer Nature.

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