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Cancer Biology

Categories
- Angiogenesis
+ Animal models
Cancer therapy
- Cell biology assays
Cell migration
+ Drug discovery and analysis
+ Cancer biochemistry
+ Cancer stem cell
+ Cell cycle checkpoints
+ Cell death
+ Cellular energetics
+ General technique
+ Genome instability & mutation
+ Inflammation
+ Invasion & metastasis
+ Microenvironment
+ Oncogenesis
+ Proliferative signaling
+ Replicative immortality
+ Tumor immunology
Protocols in Past Issues

Isolation of Murine Primary Aortic Smooth Muscle Cells
Max Ole Hubert Max Ole Hubert
Juan Rodriguez-Vita Juan Rodriguez-Vita
LW Lena Wiedmann
AF Andreas Fischer
Q&A 1 Q&A
8214 Views
Feb 5, 2021

Vascular smooth muscle cells (VSMCs) have been cultured for decades to study the role of these cells in cardiovascular disorders. The most common source of VSMCs is the rat aorta. Here we show the adaptation of this method to isolate and culture mouse aortic VSMCs. The advantage of this method is that there are many more transgenic mouse lines available compared to rats. The protocol consists of the isolation of the aorta, the liberation of vascular cells by the action of collagenase, culturing of VSCMs, and analyzing filamentous actin and alpha smooth muscle actin by fluorescence microscopy. VSCMs can be further used to study mechanisms underlying cardiovascular diseases.


Graphic abstract



Figure 1. Working steps


Human Endothelial Cell Spheroid-based Sprouting Angiogenesis Assay in Collagen
FT Fabian Tetzlaff
AF Andreas Fischer
Q&A 1 Q&A
21653 Views
Sep 5, 2018
Angiogenesis, the formation of new blood vessels from pre-existing ones plays an important role during organ development, regeneration and tumor progression. The spheroid-based sprouting assay is a well-established and robust method to study the influence of genetic alterations or pharmacological compounds on capillary-like tube formation of primary cultured endothelial cells. A major advantage of this assay is the possibility to study angiogenesis in a 3D environment. Endothelial cells are cultured as hanging drops to form spheroids. Those spheroids are embedded into a collagen matrix and tube formation is analyzed 24 h later. By analyzing sprout number and sprout length the effects of genetic manipulation or drug treatment on angiogenesis can be investigated.

In vitro Human Umbilical Vein Endothelial Cells (HUVEC) Tube-formation Assay
JK Josephine MY Ko
Maria Li Lung Maria Li Lung
Q&A 1 Q&A
70620 Views
Sep 20, 2012
Angiogenesis is involved not only in pathological conditions including cancer biology and non-neoplastic diseases, but also many biological processes including reproduction, development and repair. During angiogenesis, endothelial cells (ECs) undergo activation after binding of angiogenic factors to their receptors, release of proteases to dissolve the basement membrane, migration towards an angiogenic signal, proliferation, and an increase in cell number for new blood vessel formation. Finally, reorganization of ECs forms the three-dimensional vasculature. HUVEC tube-formation assay is one of the simple, but well-established in vitro angiogenesis assays based on the ability of ECs to form three-dimensional capillary-like tubular structures, when cultured on a gel of growth factor-reduced basement membrane extracts. During the assay, ECs differentiate, directionally migrate to align, branch, and form the tubular polygonal networks of blood vessels.