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Three-dimensional (3D) assembly of microstructures encapsulating co-cultured multiple cells can highly recapitulate the in vivo tissues, which has a great prospect in tissue engineering and regenerative medicine. In order to fully mimic the in vivo architecture, the hydrogel microstructure needs to be designed into a special shape and spatially organized without damage, which is very challenging because of its limited mechanical properties. Here, we propose a 3D assembly method for the construction of liver lobule-like microstructures (a mimetic gear-like microstructure of liver lobule) through the local fluidic interaction. Although the method has been proven and is known as the consensual means for constructing 3D cellular models, it is still challenging to improve the assembly efficiency and the assembly success rate by adjusting the fluidic force of non-contact lifting and stacking. To improve the assembly efficiency and the assembly success rate, a fluidic simulation model is proposed based on the mechanism of the interaction between the microstructures and the fluid. By computing the simulation model, we found three main parameters that affect the assembly process; they are the velocity of the microflow, the tilt angle of the manipulator and the spacing between the microstructures and the manipulator. Compared with our previous work, the assembly efficiency was significantly improved 63.8% by using the optimized parameters of the model for assembly process, and the assembly success rate was improved from 98% to 99.5%. With the assistance of the assembly simulation, the luminal 3D micromodels of liver tissue show suitable bioactivity and biocompatibility after long-term hepatocytes culture. We anticipate that our method will be capable of improving the efficiency of the microstructures assembly to regenerate more complex multicellular constructs with unprecedented possibilities for future tissue engineering applications.

micromachines

Optimization of the Fluidic-Based Assembly for Three-Dimensional Construction of Multicellular Hydrogel Micro-Architecture in Mimicking Hepatic Lobule-like Tissues