In vivo Matrigel Plug Angiogenesis Assay

The matrigel plug angiogenesis assay is a simple in vivo technique to detect the newly formed blood vessels in the transplanted gel plugs in nude mice. The matrigel matrix is derived from the engelbroth-holm-swarm (EHS) mouse sarcoma, and its composition is comparable to the basement membrane proteins. The matrigel can induce differentiation of a variety of cell types such as hepatocytes, mammary epithelial cells, and endothelial cells. In our case, tumor cells are mixed with the matrigel gel and are injected into the mice. The later immunohistochemistry (IHC) staining with the endothelial marker indicates the presence of the newly formed capillaries in the sectioned gel plugs.

Keywords: In vivo, Matrigel plug, Angiogenesis, Nude mice, Endothelial cell

Materials and Reagents

  1. BALB/cAnN-nu (Nude) female mouse, 6 to 8 week-old
  2. Tumor cells
  3. Matrigel matrix (BD Biosciences, Falcon®, catalog number: 354234 )
  4. 10% formalin solution-neutral buffered (Sigma-Aldrich, catalog number: HT501128-4L )
  5. Anti-rat CD34 (Santa Cruz, catalog number: sc-18917 )
  6. Biotin goat Anti-Rat IgG (BD Biosciences, catalog number: 559286 )
  7. Streptavidin-peroxidase conjugate (Dako, catalog number: P0397 )
  8. Diaminobenzidine (DAB) (Dako, catalog number: K3467 )
  9. Paraffin
  10. Potassium chloride
  11. Potassium phosphate monobasic
  12. Dodium chloride (NaCl)
  13. Sodium phosphate dibasic anhydrous
  14. Hematoxylin and eosin (H&E)
  15. Trypsin
  16. Complete medium
  17. Plain medium
  18. Phosphate buffered saline (PBS) (see Recipes)

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