Generation and metabolism of liver cancer organoids in alginate-chitosan hybrid microcapsules

Liver cancer had the fastest increasing mortality for decades. Compared to 2D models, that cannot simulate the microenvironment, and key functions of organs, 3D organoids models are now entering the field of in vitro analysis. Here, alginate-chitosan hybrid microcapsules are fabricated with a high reproducible cross junction based microfluidic droplet generation system. With high reproducibility and compartmentalization, 103 of microcapsules can be generated within a few minutes. The effects of fluid flow rates on its sizes and shell thicknesses were systematically studied, forming microcapsules with different capsule diameters (~ 300 to 700 µm) and thicknesses (~ 5 to 150 µm). The semi-permeability of these capsules has been well studied, combined a COMSOL model. These microcapsules, with a suitable diffusion rate for nutrients, are applied for human hepatoma cell line encapsulation. Metabolic of organoids in capsules are confirmed by tracking substrates (testosterone) and metabolites (androstenedione). Overall, highly repeatable alginate-chitosan hybrid microcapsules obtained by microfluidic droplet method are not only suitable for liver cancer cells culturing, but also promising for various cell organoids generation.