Vascular aging and dysfunction are significant contributors to age-related cardiovascular and neurodegenerative diseases. In particular, aging impacts small vessels, damaging vascular integrity leading to leakage events and inflammation, which can be further exacerbated by environmental factors. Here, we generate and evaluate an isogenic endothelial cell and pericyte venule-like co-culture model of microvasculature under perfusion with male aged human serum over 4 days. Using this model in comparison to male young human serum perfusion controls, we define the molecular and functional changes induced by aging-related circulatory cues, including functional loss of paracellular barrier integrity and modulation of transport of low-density lipoprotein. Additionally, in comparison with endothelial monoculture, we identify critical changes to basement membrane composition and aged-serum-mediated cell cycle shifts with pericyte co-culture. This modular approach reveals key impacts to further our understanding of vascular aging and to leverage in designing therapeutic and preventative approaches.