Mammary microvessels are sensitive to menstrual cycle sex hormones

Abstract

The mammary gland is a highly vascularized organ that is influenced by sex hormones including estrogen (E2) and progesterone (P4). Beyond whole-organism studies in rodents or 2D monocultures, hormonal interactions and their effects on the breast microvasculature remains largely understudied. Recent methods to generate 3D microvessels on-chip have enabled direct observation of complex vascular processes; however, these models often use non-tissue-specific cell types, such as HUVEC and fibroblasts from various sources. Here, novel mammary-specific microvessels are generated by co-culturing primary breast endothelial cells and fibroblasts under optimized culture conditions. These microvessels are mechano-sensitive (to interstitial flow) and require endothelial-stromal interactions to develop fully perfusable vessels. These mammary-specific microvessels are also responsive to exogenous stimulation by sex hormones. When treated with combined E2 and P4, corresponding to the four phases of the menstrual cycle (period, follicular, ovular, and luteal), vascular remodeling and barrier function are altered in a phase-dependent manner. The presence of high E2 (ovulation) promotes vascular growth and remodeling, corresponding to high depletion of proangiogenic factors, whereas high P4 concentrations (luteal) promote vascular regression. The effects of combined E2 and P4 hormones are not only dose-dependent but also tissue-specific, as is shown by similarly treating non-tissue-specific HUVEC microvessels.