Background
Inhibition of angiogenesis reverses rodent obesity. A validated assay in human fat tissue is needed to study the role of angiogenesis in human obesity.
Methods
Human fat tissue fragments from surgery were placed in 96-well plates, embedded in fibrin thrombin clot and overlaid with cell culture media containing 20% fetal bovine serum. After 15 days, the clots were examined by histology and electron microscopy. The effect of taxol, cobalt chloride and a heparin-steroid combination was tested in the fat tissue assay and compared to the validated human placental vein angiogenesis model (HPVAM).
Results
Blood vessels initiated growth and elongated from the fat tissue fragments over 15 days. Presence of blood vessels was confirmed with histology and electron microscopy. Taxol at 10−6 and 10−7 M completely inhibited angiogenesis, while Taxol 10−8 and 10−9 M and the heparin-steroid partially inhibited angiogenesis.The response to taxol and heparin-steroid was similar to that of the HPVAM, a validated angiogenesis assay. Cobalt chloride, a stimulator of vascular endothelial growth factor (VEGF) stimulated angiogenesis initiation at 10−9 M in fat tissue and the HPVAM, but at 10−10 M blood vessel growth was stimulated only in the fat assay.
Conclusion
This angiogenesis assay based on human fat tissue uses three-dimensionally intact human tissue. The vessels are derived from quiescient vessels within the fat. These properties allow the angiogenic switch to be evaluated in an in vitro setting. The angiogenic response of fat tissue is not identical to placental tissue. This assay allows exploration of angiogenesis in fat tissue.
Similar content being viewed by others
References
Rupnick MA, Panigrahy D, Zhang CY et al. Adipose tissue mass can be regulated through the vasculature. Proc Natl Acad Sci USA 2002; 99: 10730–. Epub 2002 Jul 29.
Gimbrone MA Jr, Cotran RS, Leapman SB et al. Tumor growth and neovascularization: an experimental model using the rabbit cornea. J Natl Cancer Inst 1974; 52: 413–7.
Ausprunk DH, Knighton DR, Folkman J. Vascularization of normal and neoplastic tissues grafted to the chick chorioallantois. Role of host and preexisting graft blood vessels. Am J Pathol 1975; 79: 597–28.
Brown KJ, Maynes SF, Bezos A et al. A novel in vitro assay for human angiogenesis. Lab Invest 1996; 75: 539–5.
Gulec SA, Woltering EA. A new in vitro assay for human tumor angiogenesis: three-dimensional human tumor angiogenesis assay. Ann Surg Oncol 2004; 11: 99–04.
Woltering EA, Lewis JM, Maxwell PJ 4th et al. Development of a novel in vitro human tissue-based angiogenesis assay to evaluate the effect of antiangiogenic drugs. Ann Surg 2003; 237: 790–; discussion 798–00.
Hornick C, Myers A, Sadowska-Krowicka H et al. Inhibition of angiogenic initiation and disruption of newly established human vascular networks by juice of Morinda Citrifola (noni). Angiogenesis 2004; 6: 143–.
Dordunoo SK, Jackson JK, Arsenault LA et al. Taxol encapsulation in poly(epsilon-caprolactone) microspheres. Cancer Chemother Pharmacol 1995; 36: 279–2.
Goldberg MA, Schneider TJ. Similarities between the oxygen-sensing mechanisms regulating the expression of vascular endothelial growth factor and erythropoietin. J Biol Chem 1994; 269: 4355–.
Jung SP, Siegrist B, Wade MR et al. Inhibition of human angiogenesis with heparin and hydrocortisone. Angiogenesis 2001; 4: 175–6.
Watson JC, Redmann JG, Meyers MO et al. Breast cancer increases initiation of angiogenesis without accelerating neovessel growth rate. Surgery 1997; 122: 508–3; discussion 513–.
Jung SP, Siegrist B, Wang YZ et al. Effect of human Angiostatin protein on human angiogenesis in vitro. Angiogenesis 2003; 6: 233–0.
Beerepoot LV, Witteveen EO, Groenewegen G et al. Recombinant human angiostatin by twice-daily subcutaneous injection in advanced cancer: a pharmacokinetic and long-term safety study. Clin Cancer Res 2003; 9: 4025–3.
Magovern CJ, Mack CA, Zhang J et al. Regional angiogenesis induced in nonischemic tissue by an adenoviral vector expressing vascular endothelial growth factor. Hum Gene Ther 1997; 8: 215–7.
Miyazawa-Hoshimoto S, Takahashi K, Bujo H et al. Elevated serum vascular endothelial growth factor is associated with visceral fat accumulation in human obese subjects. Diabetologia 2003; 46: 1483–. Epub 2003 Oct 8.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Greenway, F.L., Liu, Z., Yu, Y. et al. An Assay to Measure Angiogenesis in Human Fat Tissue. OBES SURG 17, 510–515 (2007). https://doi.org/10.1007/s11695-007-9089-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11695-007-9089-z