Skip to main content

Studying Angiogenesis Using Matrigel In Vitro and In Vivo

  • Protocol
  • First Online:
Vascular Hyperpermeability

Abstract

Angiogenesis plays a critical role in physiology and pathophysiology of the human body; hence, it is important to explore the methods to study angiogenesis under in vitro and in vivo settings. Here, we describe three different methods to assess angiogenesis using Matrigel: an in vitro two- or three-dimensional (2D/3D) tube formation or angiogenesis assay using endothelial cells with growth factor supplemented Matrigel, an ex vivo sprouting angiogenesis assay embedding aortic rings in the Matrigel, and finally, Matrigel plug assays wherein Matrigels are implanted into the flanks of mice to assess the recruitment of endothelial cells to form new blood vessels in vivo.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 139.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 179.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Carmeliet P, Jain RK (2011a) Molecular mechanisms and clinical applications of angiogenesis. Nature 473(7347):298–307

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Carmeliet P, Jain RK (2011b) Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases. Nat Rev Drug Discov 10(6):417–427

    Article  CAS  PubMed  Google Scholar 

  3. Dvorak HF, Brown LF, Detmar M, Dvorak AM (1995) Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am J Pathol 146(5):1029–1039

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Hoeben A, Landuyt B, Highley MS, Wildiers H, Van Oosterom AT, De Bruijn EA (2004) Vascular endothelial growth factor and angiogenesis. Pharmacol Rev 56(4):549–580. https://doi.org/10.1124/pr.56.4.3

    Article  CAS  PubMed  Google Scholar 

  5. Baker M, Robinson SD, Lechertier T, Barber PR, Tavora B, D’Amico G, Jones DT, Vojnovic B, Hodivala-Dilke K (2011) Use of the mouse aortic ring assay to study angiogenesis. Nat Protoc 7(1):89–104. https://doi.org/10.1038/nprot.2011.435

    Article  CAS  PubMed  Google Scholar 

  6. Malinda KM (2009) In vivo matrigel migration and angiogenesis assay. Methods Mol Biol 467:287–294. https://doi.org/10.1007/978-1-59745-241-0_17

    Article  CAS  PubMed  Google Scholar 

  7. Kruger EA, Figg WD (2001) Protein binding alters the activity of suramin, carboxyamidotriazole, and UCN-01 in an ex vivo rat aortic ring angiogenesis assay. Clin Cancer Res 7(7):1867–1872

    CAS  PubMed  Google Scholar 

  8. Devy L, Blacher S, Grignet-Debrus C, Bajou K, Masson V, Gerard RD, Gils A, Carmeliet G, Carmeliet P, Declerck PJ, Noel A, Foidart JM (2002) The pro- or antiangiogenic effect of plasminogen activator inhibitor 1 is dose dependent. FASEB J 16(2):147–154. https://doi.org/10.1096/fj.01-0552com

    Article  CAS  PubMed  Google Scholar 

  9. Cappelli HC, Kanugula AK, Adapala RK, Amin V, Sharma P, Midha P, Paruchuri S, Thodeti CK (2019) Mechanosensitive TRPV4 channels stabilize VE-cadherin junctions to regulate tumor vascular integrity and metastasis. Cancer Lett 442:15–20. https://doi.org/10.1016/j.canlet.2018.07.042

    Article  CAS  PubMed  Google Scholar 

  10. Adapala RK, Thoppil RJ, Ghosh K, Cappelli HC, Dudley AC, Paruchuri S, Keshamouni V, Klagsbrun M, Meszaros JG, Chilian WM, Ingber DE, Thodeti CK (2016) Activation of mechanosensitive ion channel TRPV4 normalizes tumor vasculature and improves cancer therapy. Oncogene 35(3):314–322. https://doi.org/10.1038/onc.2015.83

    Article  CAS  PubMed  Google Scholar 

  11. Guarino BD, Adapala RK, Kanugula AK, Lenkey NM, Dougherty JA, Paruchuri S, Khan M, Thodeti CK (2019) Extracellular vesicles from pathological microenvironment induce endothelial cell transformation and abnormal angiogenesis via modulation of TRPV4 channels. Front Cell Dev Biol 7:344. https://doi.org/10.3389/fcell.2019.00344

    Article  PubMed  PubMed Central  Google Scholar 

  12. Thoppil RJ, Cappelli HC, Adapala RK, Kanugula AK, Paruchuri S, Thodeti CK (2016) TRPV4 channels regulate tumor angiogenesis via modulation of Rho/Rho kinase pathway. Oncotarget 7(18):25849–25861. https://doi.org/10.18632/oncotarget.8405

    Article  PubMed  PubMed Central  Google Scholar 

  13. Adapala RK, Talasila PK, Bratz IN, Zhang DX, Suzuki M, Meszaros JG, Thodeti CK (2011) PKCalpha mediates acetylcholine-induced activation of TRPV4-dependent calcium influx in endothelial cells. Am J Physiol Heart Circ Physiol 301(3):H757–H765

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Dudley AC, Khan ZA, Shih SC, Kang SY, Zwaans BM, Bischoff J, Klagsbrun M (2008) Calcification of multipotent prostate tumor endothelium. Cancer Cell 14(3):201–211

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Kanugula AK, Adapala RK, Midha P, Cappelli HC, Meszaros JG, Paruchuri S, Chilian WM, Thodeti CK (2019) Novel noncanonical regulation of soluble VEGF/VEGFR2 signaling by mechanosensitive ion channel TRPV4. FASEB J 33(1):195–203. https://doi.org/10.1096/fj.201800509R

    Article  CAS  PubMed  Google Scholar 

  16. Lim YC, Garcia-Cardena G, Allport JR, Zervoglos M, Connolly AJ, Gimbrone MA Jr, Luscinskas FW (2003) Heterogeneity of endothelial cells from different organ sites in T-cell subset recruitment. Am J Pathol 162(5):1591–1601. https://doi.org/10.1016/S0002-9440(10)64293-9

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Charles K. Thodeti .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2024 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Kanugula, A.K. et al. (2024). Studying Angiogenesis Using Matrigel In Vitro and In Vivo. In: Tharakan, B. (eds) Vascular Hyperpermeability. Methods in Molecular Biology, vol 2711. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3429-5_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3429-5_9

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3428-8

  • Online ISBN: 978-1-0716-3429-5

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics