Incorporating Pericytes into an Endothelial Cell Bead Sprouting Assay

作者： Salma H. Azam1, Mitchell Smith2, Vivek Somasundaram2, Chad V. Pecot2,3 1Curriculum in Genetics and Molecular Biology,University of North Carolina at Chapel Hill, 2Lineberger Comprehensive Cancer Center,University of North Carolina at Chapel Hill, 3Department of Medicine,University of North Carolina at Chapel Hill

简介：

Overview

This protocol presents a novel in vitro bead assay that models sprouting angiogenesis by incorporating pericytes. This method allows for the study of endothelial cell and pericyte interactions critical for angiogenesis.

Key Study Components

Area of Science

• Vascular Biology
• Angiogenesis
• Cellular Interactions

Background

• Angiogenesis is a complex process involving various cell types.
• Pericytes play a crucial role in supporting endothelial cells.
• Traditional assays may not fully capture the interactions between these cell types.
• This study aims to improve in vitro models of angiogenesis.

Purpose of Study

• To develop a bead assay that incorporates pericytes.
• To better model the heterotypic interactions during angiogenesis.
• To facilitate research on endothelial cell and pericyte interactions.

Methods Used

• Reconstitution of microcarrier beads in PBS.
• Autoclave sterilization of the bead suspension.
• Aliquoting complete medium into FACS tubes.
• Adding autoclave sterilized microcarrier bead suspension and cells to tubes.

Main Results

• The assay integrates different stages of sprouting angiogenesis.
• It allows for the study of endothelial cell pericyte interactions.
• The method provides insights into the complexity of angiogenesis.
• Results can inform future vascular biology research.

Conclusions

• This novel bead assay enhances the modeling of angiogenesis.
• Incorporating pericytes is essential for accurate in vitro studies.
• The method can address key questions in vascular biology.

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