Abstract
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
Software
Equipment
Procedure
Recipes
Acknowledgments
The study was financed by the Research Grants Council of the Hong Kong Special Administrative Region, People’s Republic of China: Grant numbers HKU6617/08M and HKU6415/06M to MLL; the NIH award AR49930 to SSA; and the Swedish Cancer Society, the Swedish Research Council, the Swedish Institute, Cancer Research Institute in New York/Concern Foundation in Los Angeles and Karolinska Institute to ERZ.
References
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thanks for your message. The gel might be loss during tissue processing (paraffin embedding). That is normal.
You are right, after formalin fixation, the gel plug has to go through the serial ETOH dehydration and xylene treatment, just like preparation of other normal/tumor tissues.
Thanks for your message, Andres. How many cells did you inject? Make sure you see the tumor cells bubble after injection, if the bubble is not formed, it is most likely that the implant will disappear. Similarily, if there is no tumor cells or not enough cells in the gel plug, the impalnt will be absorbed. You can try increasing the volume of matrigel (eg. up to 450ul gel and 50ul cells) or increasing the cell numbers (up to 1 x 10e7).For the case of haematoma, have you done the tissue sectioning of the "haematoma" and subsequent H&E/CD34 staining? that happens sometimes since the tumor blood vessels are leaky.Good Luck!
Thanks a lot for your question, actually either latero or dorsal injection is acceptable.