RRC ID 38768
Author Okada N, Fushimi M, Nagata Y, Fukunaga T, Tsutsumi Y, Nakagawa S, Mayumi T.
Title A quantative in vivo method of analyzing human tumor-induced angiogenesis in mice using agarose microencapsulation and hemoglobin enzyme-linked immunosorbent assay.
Journal Jpn. J. Cancer Res.
Abstract This study was conducted to develop a quantitative assay system for use in the in vivo evaluation in mice of angiogenesis induced by human tumor cells. The human epidermoid carcinoma cells, A431 cells, were cultured on microcarriers. Microcarrier-attached A431 cells (A431-MC) were microencapsulated with agarose hydrogel to isolate them from the immune system of the C57BL/6 mice after subcutaneous dorsal midline implantation. The agarose hydrogel-microencapsulated A431 cells (Aga-A431 cells; diameter=300 micron) survived for at least 10 days in vitro, and the proliferation profile of the Aga-A431 cells was indistinguishable from that of non-microencapsulated A431 cells. The Aga-A431 cells were subcutaneously injected into mice with an 18-gauge needle. Ten days later, few vessels had formed at the site implanted with cell-free agarose beads, whereas notable angiogenesis was observed at the site implanted with Aga-A431 cells. The degree of angiogenesis was evaluated by measurement of the hemoglobin content in the implanted site using a mouse hemoglobin (mHb) enzyme-linked immunosorbent assay (ELISA) system. This mHb-ELISA system has the advantages of great simplicity and reproducibility. The measured mHb-content of new blood vessels at the site implanted with agarose beads was in good agreement with the amount of angiogenesis observed under a stereoscopic microscope. This assay system enabled us to evaluate the angiogenesis induced by xenogeneic cells, such as human tumor cells. Thus, our novel method may be useful for the study of the angiogenic potential of various human tumor cells and in research on the anti-angiogenic properties of various agents.
Volume 86(12)
Pages 1182-8
Published 1995-12
DOI 10.1111/j.1349-7006.1995.tb03313.x
PII 0910505096852604
PMID 8636008
PMC PMC5920674
MeSH Animals Antibodies, Monoclonal / immunology Carcinoma, Squamous Cell / metabolism Carcinoma, Squamous Cell / pathology* Drug Compounding Endothelial Growth Factors / physiology Endothelium, Vascular / physiopathology Enzyme-Linked Immunosorbent Assay* Fibroblast Growth Factors / physiology Hemoglobins* / administration & dosage Hemoglobins* / immunology Humans Hydrogel, Polyethylene Glycol Dimethacrylate Lymphokines / physiology Male Mice Mice, Inbred C57BL Neoplasm Proteins / physiology Neoplasm Transplantation Neovascularization, Pathologic / physiopathology* Polyethylene Glycols Rabbits Sepharose Tumor Cells, Cultured / transplantation Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factors
Times Cited 10
Human and Animal Cells