The hepatic biliary network is composed of bile canaliculi formed by hepatocytes and biliary ducts lined with cholangiocytes, connected by a heterotypic interface known as the canal of Hering. This PhD project exploits engineering approaches, such as microfabrication and micropatterning, to develop models for studying complex phenomena related to hepatocyte polarity, through two complementary projects. The first project aimed to model the hepatobiliary connection corresponding to the canal of Hering. We demonstrated the formation of heterotypic lumens (heterolumens) by confining hepatocytes and cholangiocytes in narrow microniches, which were then used for quantitative screening of different cell sources and culture media. Heterolumen formation was reproduced in a 3D model of micropatterned heterospheroids, allowing the observation of self-organization at a larger scale and confirming the observations made in microniches. Finally, a modular “spheroid-on-tube” system combining pre-formed hepatocyte spheroids and biliary tubes of defined dimensions provided a more complex model with a tunable number of hepatobiliary interfaces and polarized structures. These models constitute relevant tools for studying canal of Hering formation and hepatocyte-cholangiocyte interactions, and may be adapted for the development of organoids based on heterotypic interfaces. The second project focused on a model for aligning bile canaliculi through micropatterning hepatocytes in lines. This approach organizes cells in two-cell-wide rows, promoting elongation of bile canaliculi along the pattern axis. Lines of 25–50 µm wide induce canalicular alignment in primary hepatocytes and, in a proliferative context, in the CAN10 cell line. This model provides a robust platform for studying canalicular morphogenesis and lumen orientation under controlled geometric constraints. Together, these studies demonstrate the relevance of engineering approaches for modelling hepatocyte polarity. By combining precise spatial control with quantitative analysis, the developed systems offer a complementary alternative to organoids, combining experimental accessibility with the modelling of complex phenomena. Their modularity paves the way for future extensions, both in terms of cell type diversity and pathological applications.
Supervision: Aurélien DENIAUD (LCBM/METEOR) et Pascale DUPUIS-WILLIAMS (U1193 INSERM)
Co-supervision: Mireille Chevallet (LCBM/METEOR)