Osteosarcoma cells enhance angiogenesis visualized by color-coded imaging in the in vivo Gelfoam® assay.

Abstract

We previously described a color-coded imaging model that can quantify the length of nascent blood vessels using Gelfoam® implanted in nestin-driven green fluorescent protein (ND-GFP) nude mice. In ND-GFP mice, nascent blood vessels are labeled with GFP. We report here that osteosarcoma cells promote angiogenesis in the Gelfoam® angiogenesis assay in ND-GFP mice. Gelfoam® was initially transplanted subcutaneously in the flank of transgenic ND-GFP nude mice. Seven days after transplantation of Gelfoam®, skin flaps were made and human 143B osteosarcoma cells expressing green fluorescent protein (GFP) in the nucleus and red fluorescent protein (RFP) in cytoplasm were injected into the transplanted Gelfoam®. The control-group mice had only implanted Gelfoam®. Skin flaps were made at days 14, 21, and 28 after transplantation of the Gelfoam® to allow imaging of vascularization in the Gelfoam® using a variable-magnification small animal imaging system and confocal fluorescence microscopy. ND-GFP expressing nascent blood vessels penetrated and spread into the Gelfoam® in a time-dependent manner in both control and osteosarcoma-implanted mice. ND-GFP expressing blood vessels in the Gelfoam® of the osteosarcoma-implanted mice were associated with the cancer cells and larger and longer than in the Gelfoam®-only implanted mice (P < 0.01). The results presented in this report demonstrate strong angiogenesis induction by osteosarcoma cells and suggest this process is a potential therapeutic target for this disease.

• Authors: Fuminari Uehara, Yasunori Tome, Shinji Miwa, Yukihiko Hiroshima, Shuya Yano, Mako Yamamoto, Sumiyuki Mii, Hiroki Maehara, Michael Bouvet, Fuminori Kanaya, Robert M Hoffman
• Journal: Journal of cellular biochemistry (J Cell Biochem) Vol. 115 Issue 9 Pg. 1490-4 (Sep 2014) ISSN: 1097-4644 [Electronic] United States
• PMID: 24590470 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: © 2014 Wiley Periodicals, Inc.

Chemical References

• Luminescent Proteins
• red fluorescent protein
• Green Fluorescent Proteins

Topics

• Animals
• Cell Line, Tumor
• Female
• Gelatin Sponge, Absorbable
• Green Fluorescent Proteins (biosynthesis, genetics)
• Humans
• Implants, Experimental
• Luminescent Proteins (biosynthesis, genetics)
• Mice
• Mice, Nude
• Mice, Transgenic
• Microscopy, Confocal
• Neoplasm Transplantation
• Neovascularization, Pathologic (metabolism)
• Osteosarcoma (blood supply)