Human Endothelial Cell Spheroid-based Sprouting Angiogenesis Assay in Collagen
Commonly used acronym: In vitro sprouting assay
Scope of the method
- Human health
- Basic Research
- In vitro - Ex vivo
- Human derived cells / tissues / organs
- Endothelial cells
- growth factor supplementation
- vascular development
- blood vessel
- tumor stroma
- sprouting angiogenesis
This assay evaluates the sprouting ability of endothelial cells in a collagen matrix and it used as an in vitro model to study the formation of new blood/lymphatic vessels. The effect of pro-angiogenic growth factors or co-cultured cells can be measured by quantifying the amount of vascular sprouts that form on endothelial spheroids. Endothelial spheroids are obtained by growing endothelial cells in hanging drops, which forces the cells to adhere to each other. After generation of the spheroids, they are embedded in a collagen matrix in which endothelial growth factors or specific cell types can be embedded. Finally the amount of endothelial sprouts is quantified as a measure of the endothelial sprouting propensity.
- - Biosafety cabinet,
- - CO2 incubator,
- - Microscope.
- History of use
- Internally validated
- Published in peer reviewed journal
Pros, cons & Future potential
The method is simple and the vascular sprouts share multiple morphological characteristics of vascular tip cells in vivo.
The model is limited to evaluating sprout propensity, which is only the first step in the angiogenic cascade. The subsequent steps of tubule and network formation cannot be evaluated.
This method could also be used using conditioned media from other cell types (for example cancer cells). As such, it could be used to asses the amount of (lymph)angiogenic signaling in the secreted medium.
References, associated documents and other information
- Meçe, O., Houbaert, D., Sassano, ML. et al. Lipid droplet degradation by autophagy connects mitochondria metabolism to Prox1-driven expression of lymphatic genes and lymphangiogenesis. Nat Commun 13, 2760 (2022). https://doi.org/10.1038/s41467-022-30490-6
Contact personDiede Houbaert
OrganisationsCellular and Molecular Medicine - KU Leuven
Department of Cellular and Molecular Medicine
Laboratory of Cell Death Research & Therapy (VIB-KU Leuven)