In Vitro Cellular & Developmental Biology - Animal

, Volume 32, Issue 7, pp 409–419

Angiogenic potential of microvessel fragments established in three-dimensional collagen gels

Authors

  • James B. Hoying
    • Section of Surgical Research, Department of SurgeryUniversity of Arizona Health Sciences Center
  • Carl A. Boswell
    • Section of Surgical Research, Department of SurgeryUniversity of Arizona Health Sciences Center
  • Stuart K. Williams
    • Section of Surgical Research, Department of SurgeryUniversity of Arizona Health Sciences Center
Cellular Models

DOI: 10.1007/BF02723003

Cite this article as:
Hoying, J.B., Boswell, C.A. & Williams, S.K. In Vitro Cell.Dev.Biol.-Animal (1996) 32: 409. doi:10.1007/BF02723003

Summary

During angiogenesis, the microvasculature displays both vessel remodeling and expansion under the control of both cellular and extracellular influences. We have evaluated the role of angiogenic and angiostatic molecules on angiogenesis in anin vitro model that more appropriately duplicates the cellular and extracellular components of this process. Freshly isolated microvessel fragments from rat adipose tissue (RFMF) were cultured within three-dimensional collagen I gels. These fragments were characterized at the time of isolation and were composed of vessel segments observed in the microvasculature of fatin situ (i.e., arterioles, venules, and capillaries). Fragments also exhibited characteristic ablumenally associated cells including smooth muscle cells and pericytes. Finally, fragments were encased in an extracellular matrix composed of collagen type IV and collagen type I/III. The elongation of microvascular elements was subsequently evaluated using morphologic and immunocytochemical techniques. The proliferation, migration, and elongation of cellular elements in microvessel fragments from rat adipose tissue was dependent on initial fragment density, matrix density, and required serum. Inclusion of endothelial cell growth factors to microvessel fragments from rat adipose tissue 3-D cultures resulted in the accelerated elongation of tube structures and the expression of von Willebrand factor in cells constituting these tubes. Molecules with reported angiostatic capacity (e.g., transforming growth factor and hydrocortisone) inhibited vessel tube elongation.

In vitro methods have been developed to evaluate numerous mechanisms associated with angiogenesis, including endothelial cell proliferation, migration, and phenotypic modulation. Microvascular endothelial cell fragments described in this study represent anin vitro population of cells that accurately duplicate thein vivo microcirculatory elements of fat. The proliferation of cells and elongation of microvascular elements subsequently observed in three-dimensional cultures provides anin vitro model of angiogenesis. Microvascular formation in this system results from pre-existing microvessel fragments unlike tube formation observed when cultured endothelial cells are placed in three-dimensional gels. This form of tube formation from cultured endothelium is more characteristic of vasculogenesis. Thus, the formation of microvascular elements from microvessel fragments provides the opportunity to examine the mechanisms regulating angiogenesis in anin vitro system amenable to precise experimental manipulation.

Key words

angiogenesisendotheliumpericyteextracellular matrixgrowth factors

Copyright information

© Society for In Vitro Biology 1996