Biology subjects

Vulto, P.

Publications and source records attributed to Vulto, P..

5 recordsLinked to original sources

bioRxiv2026

Scalable microvascular networks-on-chip enable 8-week unidirectional perfusion for long-term vascular toxicity screening

Cardiovascular toxicity is a leading cause of late-stage drug attrition, yet current in vitro vascular models lack the longevity and scalability needed to capture clinically relevant responses in mature vasculature. We developed a vascular microphysiological system comprising 32 parallel, self-assembled microvascular networks from primary human endothelial cells and stromal fibroblasts. Networks were unidirectionally perfused by gravity-driven, pump-free flow. They remained functionally perfusable for at least 57 days, the longest duration reported for self-assembled microvascular networks, and underwent progressive maturation characterised by perivascular fibroblast organisation, basement membrane deposition, and matrix remodelling. Variance decomposition confirmed high reproducibility, with intra-plate variability of 3-7% and no operator-dependent effects on network morphometry. Exploiting this extended culture window, we reveal maturation-dependent shifts in endothelial inflammatory responsiveness and cytotoxic susceptibility. Distinct acute and chronic toxicity profiles of clinically used tyrosine kinase inhibitors are resolved, and a concentration- and time-dependent spectrum of sorafenib-induced microvascular responses is produced. These results show that vascular longevity and progressive maturation are key requirements for assessing vascular toxicity, and provide a scalable platform for evaluating acute and chronic drug responses in mature human microvascular tissues.

cell biology↗

bioRxiv2025

Modeling Hepatocellular Carcinoma and its microenvironment on a chip

Hepatocellular carcinoma (HCC) is the most common type of liver cancer. Its incidence is increasing and is closely related to advanced liver disease. Interactions in the HCC microenvironment between tumor cells and the associated stroma actively regulate tumor initiation, progression, metastasis, and therapy response. Effective drug development increasingly requires advanced models that can be utilized in the earliest stages of compound and target discovery. Here we report a phenotypic screen on an advanced HCC patient-derived chip (PDChip) model. The vascularized HCC PDChip models include relevant cellular players of the HCC microenvironment. We assessed the effect of 28 treatment conditions on a panel of 8 primary HCC tumors and 2 cell lines. Approximately 1200 HCC PDchips were grown under perfusion flow, exposed to treatments, and subsequently assessed for viability, tumor-associated vasculature responses and chemokine and cytokine changes. Although the SoC therapeutics sorafenib and lenvatinib reduced culture viability and produced profound changes in the organization of the vascular beds, they did not affect the tumor cell population in these cultures. Atorvastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor, reduced tumor viability but did not affect vascular bed organization. Sorafenib, lenvatinib and atorvastatin also affected chemokine and cytokine release. Tocilizumab, galunisertib, and vactosertib decreased the level of IL6, a relevant prognostic marker for HCC, while IL6 was increased by halofuginone. In conclusion, HCC PDChip models enabled a detailed evaluation of drug-induced responses in the tumor and associated microenvironment, highlighting their importance in preclinical research for understanding diseases and developing new drugs.

bioengineering↗

bioRxiv2024

A microvascularized in vitro liver model for disease modeling and drug discovery

Drug discovery for complex liver diseases faces alarming attrition rates. The lack of non-clinical models that recapitulate key aspects of liver (patho)-physiology is likely contributing to the inefficiency of developing effective treatments. Of particular notice is the common omission of an organized microvascular component despite its importance in maintaining liver function and its involvement in the development of several pathologies. Increasing the complexity of in vitro models is usually associated with a lack of scalability and robustness which hinders their implementation in drug development pipelines. Here, we describe a comprehensive liver MPS model comprising stellates, liver-derived endothelial cells and hepatocytes conceived within a scalable and automated platform. We show that endothelial cells self-organize in a microvascular network when co-cultured with stellates in a hydrogel. In a tri-culture, hepatocytes polarize accordingly, with a basolateral side facing blood vessels and an apical side facing bile-canaliculi-like structures. Stellates interact and surround the hollow microvessels. Steatosis was induced by exogenous administration of fatty acids which could be prevented by co-administration of firsocostat. Administration of TGF-{beta} resulted in an activated stellate cells phenotype which could be prevented by the co-administration of SB-431542. The model was implemented on a microtiter plate format comprising 64 chips which enabled the development of a fully automated, multiplexed fibrosis assay with a robust Z factor suitable for high-throughput applications.

bioengineering↗

bioRxiv2022

Healthy and diseased placenta barrier on-a-chip models suitable for high-throughput studies

Preeclampsia is one of the leading causes of maternal and perinatal morbidity in the world. Its fundamental mechanisms are yet poorly understood due to a lack of good experimental models. Here we report an in vitro model of the placental barrier, based on co-culture of trophoblasts and endothelial cells against a collagen extracellular matrix in a microfluidic platform. The model yielded a functional syncytium with barrier properties, polarization, secretion of relevant extracellular membrane components, thinning of the materno-fetal space, hormone secretion, and transporter function. The model was exposed to low oxygen conditions and perfusion flow was modulated to induce preeclamptic characteristics, resulting in reduced barrier function, hormone secretion, and microvilli as well as increased nuclei count. The model was implemented in a titer plate microfluidic platform fully amenable to high-throughput screening. We thus believe this model could aid mechanistic understanding of preeclampsia as well as support future development of effective therapies through target and compound screening campaigns.

bioengineering↗

bioRxiv2017

Automated microuidic cell culture of stem cell derived dopaminergic neurons in Parkinson’s disease

Parkinsons disease is a slowly progressive neurodegenerative disease characterised by dysfunction and death of selectively vulnerable midbrain dopaminergic neurons leading mainly to motor dysfunction, but also other non-motor symptoms. The development of human in vitro cellular models with similar phenotypic characteristics to selectively vulnerable neurons is a major challenge in Parkinsons disease research. We constructed a fully automated cell culture platform optimised for long-term maintenance and monitoring of induced pluripotent stem cell derived neurons in three dimensional microfluidic cell culture devices. The system can be flexibly adapted to various experimental protocols and features time-lapse imaging microscopy for quality control and electrophysiology monitoring to assess neuronal activity. Using this system, we continuously monitored the differentiation of Parkinsons disease patient derived human neuroepithelial stem cells into midbrain specific dopaminergic neurons. Calcium imaging confirmed the electrophysiological activity of differentiated neurons and immunostaining confirmed the efficiency of the differentiation protocol. This system is the first example of a fully automated Organ-on-a-Chip culture and enables a versatile array of in vitro experiments for patient-specific disease modelling.

bioengineering↗