Microtissue Geometry and Cell‐Generated Forces Drive Patterning of Liver Progenitor Cell Differentiation in 3D

Abstract

3D microenvironments provide a unique opportunity to investigate the impact of intrinsic mechanical signaling on progenitor cell differentiation. Using a hydrogel‐based microwell platform, arrays of 3D, multicellular microtissues in constrained geometries, including toroids and cylinders are produced. These generated distinct mechanical profiles to investigate the impact of geometry and stress on early liver progenitor cell fate using a model liver development system. Image segmentation allows the tracking of individual cell fate and the characterization of distinct patterning of hepatocytic makers to the outer shell of the microtissues, and the exclusion from the inner diameter surface of the toroids. Biliary markers are distributed throughout the interior regions of micropatterned tissues and are increased in toroidal tissues when compared with those in cylindrical tissues. Finite element models of predicted stress distributions, combined with mechanical measurements, demonstrates that intercellular tension correlates with increased hepatocytic fate, while compression correlates with decreased hepatocytic and increased biliary fate. This system, which integrates microfabrication, imaging, mechanical modeling, and quantitative analysis, demonstrates how microtissue geometry can drive patterning of mechanical stresses that regulate cell differentiation trajectories. This approach may serve as a platform for further investigation of signaling mechanisms in the liver and other developmental systems.

Stereo Coverage and Overall Stiffness of Biomaterial Arrays Underly Parts of Topography Effects on Cell Adhesion

“…Surface patterning serves as a fundamental tool to explore the principle of cell–material interactions . These corresponding techniques are divided into chemical patterning − and physical patterning. , Based on well-designed surface patterns, researchers can adjust topology, stiffness, and external mechanical force; and the patterning effect can be combined with external physical stimuli, degradation rate, and so forth.…”

Spatially Defined Cell-Secreted Protein Detection Using Granular Hydrogels: μGeLISA

“…The microwell composition and production process have been described previously . 4-arm PEG acrylate was dissolved to 10% w/v [in LAP-PBS.…”

“…The microwell composition and production process have been described previously. 39 4-arm PEG acrylate was dissolved to 10% w/v [in LAP-PBS. 0.5 μL of the PEG acrylate solution was placed on the well and flattened with an 8 mm diameter coverslip and exposed to 67.62 mW/cm of UV light for 30 s to form a thin ∼10 μm PEG layer to prevent cell adhesion to the glass.…”

Control of hydrostatic pressure and osmotic stress in 3D cell culture for mechanobiological studies

“…Integrating this basis with recent breakthroughs in tissue fabrication, biophysics, and biomaterials, should enable bottom-up strategies for the design of bioreactors that are well-suited to investigate the mechanobiology of HP and OS. Novel methods in 3D tissue bioprinting [159][160][161][162][163], mechanical characterization of biological tissues [164][165][166][167][168][169], the formulation of responsive biomaterials with tunable biomechanical and biochemical properties [170][171][172][173][174][175][176], and the development of 3D organotypic cell cultures [177][178][179][180][181][182] highlight a battery of innovations that can be exploited towards the design of advanced materials systems and bioreactors for the control of HP and OS.…”