Biology subjects

Mazari-Arrighi, E.

Publications and source records attributed to Mazari-Arrighi, E..

5 recordsLinked to original sources

bioRxiv2026

Biomimetic 3D mammary duct models of healthy and tumoral tissues engineered by a co-extrusion microfluidic based technology

Engineering the human breast in 3D physio-mimetic models is challenging due to its complex multilayered tubular organization, where milk is produced in acini and transported through ductal structures. These functions rely on a highly organized architecture comprising stromal, epithelial, and extracellular matrix compartments. The dysregulation of this architecture perturbs mammary gland homeostasis and promotes the emergence of diverse breast cancer subtypes, from frequent in situ luminal to rarer metastatic basal-like tumors. Despite this knowledge, conventional anti-cancer drug testing still primarily employs high-throughput 3D spheroid models that account for diffusion but lack stromal components, thereby failing to capture stroma-driven treatment resistance. With a unique microfluidic co-extrusion platform, we have developed 3D tubular tissues anchored on a porous and biocompatible alginate shell. Using a one-step protocol, we have bioengineered six relevant ductoid models of healthy and tumoral mammary ducts, most notably a multi-layered model comprising a lumen, mammary epithelium, and stromal compartment made of fibroblasts and matrixes. These new models offer limitless applications in tissue engineering including the characterization of an epithelium and its secretory function, and the identification of the stromal influence on healthy and tumoral mammary gland tissue. Finally, by releasing mechanical constraints, we scale-up the tubular duct model into a mammary assembloid that exhibits branching and budding of acini-like structures from the original duct. We envision that this modular design will broadly impact breast basic and clinical research by opening new experimental avenues toward more physio-mimetic tools through the integration of stromal compartments.

bioengineering↗

bioRxiv2025

A versatile microfluidic extrusion-based hydrogel platform for self-organization and long-term maintenance of engineered 3D lymphatic endothelium

Lymphatic endothelium is essential for interstitial fluid drainage, immune surveillance, and macromolecular transport. Existing in vitro models that accurately recapitulate its three-dimensional structure, barrier properties, and long-term stability are still limited. In this study, we propose a robust, adaptable, and scalable platform designed to generate engineered three-dimensional lymphatic endothelium using primary human dermal lymphatic endothelial cells (HDLECs). This system combines microfluidic extrusion-based fabrication allowing precise control of the tubular geometry with custom-optimized extracellular matrix-derived hydrogel. The capability to tune dimensions enables the creation of constructs that encompass physiologically relevant size ranges. Through a systematic matrix screen, we identified a unique four-component formulation--gelatin, Matrigel, hyaluronic acid, and fibrinogen--that supports the rapid self-assembly of HDLECs into stable, lumen-forming monolayers within one week. These engineered structures maintain viability and structural integrity for a minimum of 30 days under static culture conditions. Functional permeability assays demonstrated selective tracer uptake characteristics of lymphatic transport. Comparative studies with blood vascular endothelial cells indicate that the proposed platform preserves maintenance of lineage-specific expression profiles, junctional organization, and permeability properties under identical fabrication parameters. Altogether, this approach offers a reproducible and controlled system for studying three-dimensional endothelial architecture, transport mechanisms, and extracellular matrix remodeling across diverse endothelial phenotypes, thereby addressing a pivotal gap in the modeling of lymphatic vasculature. HighlightsO_LIA microfluidic extrusion platform allows primary human lymphatic endothelial cells to self-organize into 3D hollow tubes. C_LIO_LIThe internal dimensions of the hybrid cell/hydrogel constructs can be precisely controlled by tuning the flow rates. C_LIO_LIA custom-defined four-component extracellular matrix provides essential biochemical cues for stable monolayer formation and for the in vitro organization of lymphatic endothelial vessels. C_LIO_LIMatrix composition significantly influences cell proliferation and migration, thereby impacting on the spatial arrangement of the tissue. C_LIO_LIThe three-dimensional lymphatic tissue configuration sustains cell viability, consistent expression of lineage-specific markers, and barrier function which lasts for at least 30 days. C_LIO_LIThe system maintains morphological and functional properties not only for lymphatic endothelial cells, but also for blood endothelial cells in long-term culture conditions. C_LI

bioengineering↗

bioRxiv2023

Self-organization of Long-lasting Human Endothelial Capillary Networks guided by DLP Bioprinting

Tissue engineering holds great promise for regenerative medicine, drug discovery and as an alternative to animal models. However, as soon as the dimensions of engineered tissue exceed the diffusion limit of oxygen and nutriments, a necrotic core forms leading to irreversible damage. To overcome this constraint, the establishment of a functional perfusion network is essential and is a major challenge to be met. In this work, we explore a promising Digital Light Processing (DLP) bioprinting approach to encapsulate endothelial progenitor cells (EPCs) in 3D photopolymerized hydrogel scaffolds to guide them towards vascular network formation. We observed that EPCs encapsulated in the appropriate photopolymerized hydrogel can proliferate and self-organize within a few days into branched tubular structures with predefined geometry, forming capillary-like vascular tubes or trees of various diameters (in the range of 10 to 100 m). Presenting a monolayer wall of endothelial cells strongly connected by tight junctions around a central lumen, these structures can be microinjected with fluorescent dye and are stable for several weeks in vitro. Interestingly, our technology has proven to be versatile in promoting the formation of vascular structures using a variety of vascular cell lines, including EPCs, human vascular endothelial cells (HUVECs) and human dermal lymphatic endothelial cells (HDLECs). We have also demonstrated that these vascular structures can be recovered and manipulated in an alginate patch without altering their shape or viability. This opens new opportunities for future applications, such as stacking these endothelial vascular structures with other cell sheets or multicellular constructs to yield bioengineered tissue with higher complexity and functionality.

bioengineering↗

bioRxiv2021

In Vitro Proliferation and Long-Term Preservation of Functional Primary Rat Hepatocytes in Cell Fibers

Primary hepatocytes are essential cellular resource for drug screening and medical transplantation. Since culture systems for them have already succeeded in reconstituting the biomimetic microenvironment, acquiring additional capabilities both to expand primary hepatocytes and to handle them easily would be expected as progress to the next stage. This paper describes a culture system for primary rat hepatocytes that is equipped with scalability and handleability relying on cell fiber technology. Cell fibers are cell-laden core-shell hydrogel microfibers; in the core regions, cells are embedded in extracellular matrix proteins, cultured three-dimensionally, and exposed to soluble growth factors in the culture medium through the hydrogel shells. By encapsulating primary rat hepatocytes within cell fibers, we first demonstrated they increase in number while keeping their viability and their hepatic specific functions for up to thirty days of subsequent culture. Then, we demonstrated the potency of the primary rat hepatocytes that proliferate in cell fibers not only as cell-based sensors to detect drugs that damage hepatic functions and hepatocellular processes but also as transplants to improve the plasma albumin concentrations of congenital analbuminemia. Therefore, our culture system could serve for innovating strategies and promising developments in applying primary hepatocytes to both pharmaceutical and medical fields.

bioengineering↗

bioRxiv2021

Construction of functional biliary epithelial branched networks with predefined geometry using digital light stereolithography

Cholangiocytes, biliary epithelial cells, are known to spontaneously self-organize into spherical cysts with a central lumen. In this work, we explore a promising biocompatible stereolithographic approach to encapsulate cholangiocytes into geometrically-controlled 3D hydrogel structures to guide them towards the formation of branched tubular networks. We demonstrate that within the appropriate mix of hydrogels, normal rat cholangiocytes can proliferate, migrate and organize into branched tubular structures, form walls consisting of a cell monolayer, transport fluorescent dyes into the luminal space and show markers of epithelial maturation such as primary cilia. The resulting structures have dimensions typically found in the intralobular and intrahepatic bile ducts and are stable for weeks, without any requirement of bulk supporting material, thereby offering total access to the basal side of these biliary epithelial constructs.

bioengineering↗