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Biology subjects

Cherubini, M.

Publications and source records attributed to Cherubini, M..

3 recordsLinked to original sources

Label-free phenotyping of human microvessel networks

Understanding the spatial heterogeneity in blood vessel formation and development is crucial for various biomedical applications. Traditional methods for in-vitro microvessel segmentation rely on fluorescent labeling, which either interferes with the sample homeostasis, limits the study to a restricted set of precursor cells, or requires sample fixation, thus preventing live measurements. Moreover, these methods often focus on small, cropped images, neglecting global spatial heterogeneity of microvasculature, leading to biased data interpretation. To overcome these limitations, we present VascuMap, a deep-learning-based tool for label-free vessel segmentation and spatial analysis. VascuMap enables a comprehensive examination of entire vessel networks, capturing both morphological and topological features across the full vascular bed. Our method achieves high segmentation accuracy, comparable to the state-of-the-art fluorescence-based models. VascuMaps capabilities extend to characterizing vasculature generated from label-free patient-derived samples, a vital step towards personalized medicine. Its compatibility with widefield label-free microscopy also accelerates sample acquisition, making it ideal for high-throughput systems crucial for drug toxicity and safety screens.

bioengineering↗

Flow in fetoplacental microvessels in vitro enhances perfusion, barrier function, and matrix stability

Proper placental vascularization is vital for pregnancy outcomes, but assessing it with animal models and human explants has limitations. Here, we present a 3D in vitro model of human placenta terminal villi that includes fetal mesenchyme and vascular endothelium. By co-culturing HUVEC, placental fibroblasts, and pericytes in a macro-fluidic chip with a flow reservoir, we generate fully perfusable fetal microvessels. Pressure-driven flow is crucial for the growth and remodeling of these microvessels, resulting in early formation of interconnected placental vascular networks and maintained viability. Computational fluid dynamics simulations predict shear forces, which increase microtissue stiffness, decrease diffusivity and enhance barrier function as shear stress rises. Mass-spec analysis reveals the deposition of numerous extracellular proteins, with flow notably enhancing the expression of matrix stability regulators, proteins associated with actin dynamics, and cytoskeleton organization. Our model provides a powerful tool for deducing complex in vivo parameters, such as shear stress on developing vascularized placental tissue, and holds promise for unraveling gestational disorders related to the vasculature.

bioengineering↗

Mammary microvessels are sensitive to menstrual cycle sex hormones

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.

bioengineering↗