Lymphatic and blood microvascular systems are critical for tissue function and remodeling. New insights into the coordination of both systems can be gained by better investigating the relationships between lymphangiogenesis and angiogenesis in controlled environments. Recently, our laboratory has established that in the rat mesentery culture model lymphatic and blood microvascular networks remain intact and viable (Stapor et al., 2013). The objective of this study was to determine whether the rat mesentery culture model can be used to identify temporal relationships between lymphangiogenesis and angiogenesis induced by vascular endothelial growth factor-C (VEGF-C). Adult male Wistar rats (n=4) were anaesthetized by i.m. injection of ketamine (80 mg/kg) and xylazine (8 mg/kg) and then euthanized. Mesenteric tissue windows were aseptically harvested and cultured for 3 days or 5 days in either serum-free minimum essential media (MEM) or MEM supplemented with VEGF-C (100 ng/ml) (n=10-12 tissues per group). Tissues were immunolabeled for PECAM and LYVE-1 to identify blood and lymphatic endothelial cells, respectively. Tissues selected randomly from those containing vascular networks were quantified for angiogenesis and lymphangiogenesis (n=4-6 tissues per group). Values are means ± S.E.M. and were statistically compared using Student’s t-tests. VEGF-C treatment resulted in a significant increase in the density of blood vessel sprouting compared to MEM controls by day 3 (22.2±1.8 vs. 13.8±1.6 sprouts/mm2, p<0.01). By day 5, lymphatic sprouting was significantly increased along the hierarchy of intact lymphatic networks compared to MEM controls (7.2±0.6 vs. 4.0±0.4 sprouts/mm2, p<0.01). These results are consistent with in vivo findings that lymphangiogenesis lags angiogenesis after chronic stimulation (Sweat et al., 2012; Baluk et al., 2005) and the results establish the rat mesentery culture model as a tool to investigate the interrelationships between lymphangiogenesis and angiogenesis ex vivo in a multisystem environment.