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A quantitative assay using basement membrane extracts to study tumor angiogenesis in vivo
Author(s) -
Ito Yasumasa,
Iwamoto Yukihide,
Tanaka Kazuhiro,
Okuyama Kiyotaka,
Sugioka Yoichi
Publication year - 1996
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19960703)67:1<148::aid-ijc24>3.0.co;2-9
Subject(s) - matrigel , ht1080 , basement membrane , angiogenesis , fibrosarcoma , in vivo , in vitro , endothelial stem cell , gentamicin protection assay , microbiology and biotechnology , chemistry , biology , vascular endothelial growth factor , neovascularization , cancer research , pathology , medicine , cancer , metastasis , biochemistry , vegf receptors , genetics
We describe a quantitative assay for assessing tumor angiogenesis in vivo using basement membrane extracts (Matrigel). Nude mice were injected s.c. with liquid Matrigel mixed with HT1080 human fibrosarcoma cells. Since Matrigel rapidly forms a solid gel at body temperature, the gel containing tumor cells can be removed immediately and then processed for histological studies. Tumor angiogenesis was monitored quantitatively by measuring both the number and the total area of neovessels present in the gels using an image analyzer, which could be achieved approximately 72 hr later. Furthermore, HT1080 cell‐conditioned medium, which may contain various tumor‐derived factors, promoted the basement membrane degradation, migration, proliferation and tube formation of endothelial cells in vitro , as did Matrigel, although to a lesser extent. In addition, Northern blot analysis demonstrated that the amount of vascular endothelial growth factor (VEGF) mRNA in HT1080 cells was much higher than that in human fibroblasts or NIH3T3 cells. Our results suggest that angiogenesis observed in our assay may be due to the synergic effects of tumor angiogenic factors such as VEGF, and Matrigel. The advantages of our assay are: 1) it is possible to assess early angiogenesis quantitatively; and 2) this assay may be applicable for screening anti‐angiogenic therapeutic agents to be used against human neoplasms. © 1996 Wiley‐Liss, Inc.