An ex vivo model for anti-angiogenic drug testing on intact microvascular networks.
New models of angiogenesis that mimic the complexity of real microvascular networks are needed. Recently, our laboratory demonstrated that cultured rat mesentery tissues contain viable microvascular networks and could be used to probe pericyte-endothelial cell interactions. The objective of this study was to demonstrate the efficacy of the rat mesentery culture model for anti-angiogenic drug testing by time-lapse quantification of network growth. Mesenteric windows were harvested from adult rats, secured in place with an insert, and cultured for 3 days according to 3 experimental groups: 1) 10% serum (angiogenesis control), 2) 10% serum + sunitinib (SU11248), and 3) 10% serum + bevacizumab. Labeling with FITC conjugated BSI-lectin on Day 0 and 3 identified endothelial cells along blood and lymphatic microvascular networks. Comparison between day 0 (before) and 3 (after) in networks stimulated by 10% serum demonstrated a dramatic increase in vascular density and capillary sprouting. Growing networks contained proliferating endothelial cells and NG2+ vascular pericytes. Media supplementation with sunitinib (SU11248) or bevacizumab both inhibited the network angiogenic responses. The comparison of the same networks before and after treatment enabled the identification of tissue specific responses. Our results establish, for the first time, the ability to evaluate an anti-angiogenic drug based on time-lapse imaging on an intact microvascular network in an ex vivo scenario.
Department of Biomedical Engineering, Tulane University, New Orleans, Louisiana, United States of America.
Mohammad S
MS
Azimi
Department of Biostatistics & Bioinformatics, Tulane University, New Orleans, Louisiana, United States of America.
Leann
L
Myers
Department of Mathematics, Tulane University, New Orleans, Louisiana, United States of America.
Michelle
M
Lacey
Department of Biomedical Engineering, Tulane University, New Orleans, Louisiana, United States of America.
Scott A
SA
Stewart
Department of Biomedical Engineering, Tulane University, New Orleans, Louisiana, United States of America.
Qirong
Q
Shi
Department of Pharmacology, Tulane University, New Orleans, Louisiana, United States of America.
Prasad V
PV
Katakam
Department of Pharmacology, Tulane University, New Orleans, Louisiana, United States of America.
Debasis
D
Mondal
Department of Biomedical Engineering, Tulane University, New Orleans, Louisiana, United States of America.
Walter L
WL
Murfee
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D000068258:Bevacizumab / Q000494:pharmacology; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D008297:Male; D008638:Mesenteric Arteries / Q000166:cytology / Q000187:drug effects*; D008853:Microscopy; D055806:Microvessels / Q000187:drug effects; D008954:Models, Biological; D011758:Pyrroles / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D000077210:Sunitinib; D059008:Time-Lapse Imaging / Q000379:methods; D046509:Tissue Culture Techniques / Q000379:methods*
D020533:Angiogenesis Inhibitors; D007211:Indoles; D011758:Pyrroles; D000068258:Bevacizumab; D000077210:Sunitinib