The DOI matches its Crossref record; 35 of the 35 references it cites were found in a registry or an index.
In vitro angiogenesis: endothelial cell tube formation on gelled basement membrane extract
Abstract
A protocol is presented here for a rapid, quantitative and reliable in vitro angiogenesis assay that can be adapted for high throughput use. Endothelial cells are plated on a gelled basement matrix, their natural substrate, and form capillary-like structures with a lumen. The assay can be used to identify inhibitors or stimulators of angiogenesis, as well as genes and signaling pathways involved in angiogenesis. It has also been used to identify endothelial progenitor cells. This assay involves endothelial cell adhesion, migration, protease activity and tubule formation. This tube formation assay is preferred, as other in vitro assays for angiogenesis, such as cell adhesion, migration and invasion, measure limited steps in the angiogenesis process. The tube formation assay on basement membrane can be completed in a day because transformed endothelial cells form tubes within 3 h, whereas non-transformed endothelial cells form tubes within 6 h.
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“together, the assays robustly assess the health of ECs.”
From An inflammatory aging clock (iAge) based on deep learning tracks multimorbidity, immunosenescence, frailty and cardiovascular aging (2021) · cites this paper as [53]
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Cites
Matrigel: Basement membrane matrix with biological activity
Kleinman, H. K., & Martin, G. R. (2005) · Seminars in Cancer Biology · doi:10.1016/j.semcancer.2005.05.004 · Cited by 1,673
Self-citationThree-dimensional culture models of normal and malignant breast epithelial cells
Lee, G. Y., Kenny, P. A., Lee, E. H., & Bissell, M. J. (2007) · Nature Methods · doi:10.1038/nmeth1015 · Cited by 1,336
HMEC-1: Establishment of an Immortalized Human Microvascular Endothelial Cell Line
Ades, E. W., Candal, F. J., Swerlick, R. A., George, V. G., Summers, S., Bosse, D. C., & Lawley, T. J. (1992) · Journal of Investigative Dermatology · doi:10.1111/1523-1747.ep12613748 · Cited by 1,265
Kubota, Y., Kleinman, H. K., Martin, G. R., & Lawley, T. J. (1988) · The Journal of Cell Biology · doi:10.1083/jcb.107.4.1589 · Cited by 1,210
Self-citationMatrix metalloproteinases and angiogenesis
Rundhaug, J. E. (2005) · Journal of Cellular and Molecular Medicine · doi:10.1111/j.1582-4934.2005.tb00355.x · Cited by 978
Grant, D. S., Tashiro, K., Seguí‐Real, B., Yamada, Y., Martin, G. R., & Kleinman, H. K. (1989) · Cell · doi:10.1016/0092-8674(89)90945-8 · Cited by 740
Self-citationAngiogenesis Assays: A Critical Overview
Auerbach, R., Lewis, R. L., Shinners, B., Kubai, L., & Akhtar, N. (2003) · Clinical Chemistry · doi:10.1373/49.1.32 · Cited by 705
Vukicevic, S., Kleinman, H. K., Luyten, F. P., Roberts, A. B., Roche, N. S., & Reddi, A. H. (1992) · Experimental Cell Research · doi:10.1016/0014-4827(92)90397-q · Cited by 696
Self-citationMorales, D., McGowan, K. A., Grant, D. S., Maheshwari, S. R., Bhartiya, D., Cid, M. C., Kleinman, H. K., & Schnaper, H. W. (1995) · Circulation · doi:10.1161/01.cir.91.3.755 · Cited by 463
Self-citationArnaoutova, I., George, J., Kleinman, H. K., & Benton, G. (2009) · Angiogenesis · doi:10.1007/s10456-009-9146-4 · Cited by 454
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Three-dimensional organotypic culture: experimental models of mammalian biology and disease
Shamir, E. R., & Ewald, A. J. (2014) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3873 · Cited by 719
Sayed, N., Huang, Y., Nguyen, K., Krejciova-Rajaniemi, Z., Grawe, A. P., Gao, T., Tibshirani, R., Hastie, T., Alpert, A., Cui, L., et al. (2021) · Nature Aging · doi:10.1038/s43587-021-00082-y · Cited by 572
LRG1 promotes angiogenesis by modulating endothelial TGF-β signalling
Wang, X., Abraham, S., McKenzie, J. A. G., Jeffs, N. R., Swire, M., Tripathi, V. B., Luhmann, U. F. O., Lange, C., Zhai, Z., Arthur, H. M., et al. (2013) · Nature · doi:10.1038/nature12345 · Cited by 541
Matrigel: From discovery and ECM mimicry to assays and models for cancer research
Benton, G., Arnaoutova, I., George, J., Kleinman, H. K., & Koblinski, J. (2014) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2014.06.005 · Cited by 477
Self-citationComprehensive characterization of the Published Kinase Inhibitor Set
Elkins, J. M., Fedele, V., Szklarz, M., Abdul Azeez, K. R., Salah, E., Mikolajczyk, J., Romanov, S., Sepetov, N., Huang, X.-P., Roth, B. L., et al. (2016) · Nature Biotechnology · doi:10.1038/nbt.3374 · Cited by 323
Metadata
- Authors
- Arnaoutova, I., & Kleinman, H. K.
- Affiliations
- Trevigen (US)
- National Institutes of Health (US)
- Corresponding author
- Arnaoutova, Irina
- Kleinman, Hynda K
- ORCID
- Kleinman, Hynda K
- Venue
- Nature Protocols
- Published
- 2010-03-11
- Volume
- 5
- Issue
- 4
- Pages
- 628-635
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W2015186671
- Cited by
- 760
- References
- 35
- Citations by year
- 2026: 29 · 2025: 60 · 2024: 55 · 2023: 54 · 2022: 60 · 2021: 62 · 2020: 57 · 2019: 56 · 2018: 59 · 2017: 66 · 2016: 54 · 2015: 31 · 2014: 41 · 2013: 33 · 2012: 26
- FWCI
- 9.89
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Protocols (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Angiogenesis and VEGF in Cancer
- Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Keywords
- in vitro angiogenesis assay, endothelial tube formation, basement membrane extract, capillary-like structures, endothelial cell adhesion, endothelial cell migration, proteolytic activity, endothelial progenitor cells, angiogenesis inhibitors, high-throughput screening
- MeSH terms
- Basement Membrane, Capillaries, Cells, Cultured, Cytological Techniques, Gels, Humans, Neovascularization, Physiologic, Endothelial Cells, In Vitro Techniques
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