A high-throughput microfluidic platform for functional hiPSC-derived liver organoids with bile duct- and lobule-like structures.

HiPSC-derived organoids have attracted significant attention in stem cell research and regenerative medicine. However, obtaining functional organoids in sufficient quantities, consistent sizes, and reproducible formats remains a significant challenge. Here, we present an innovative microfluidic platform for the high-throughput production of HAMA core-shell microspheres (HCSM) for the encapsulation and differentiation of hiPSCs. Fish gelatin and HAMA were employed as the core and shell materials, respectively. Using this platform, we successfully fabricated HCSM with uniform and controllable sizes in a high-throughput manner. Single-cell hiPSC suspensions self-organized into spheroids within HCSM, leading to the formation of EBs exhibiting cavitation. These EBs effectively differentiated into brain organoids, beating cardiac organoids, and liver organoids. Detailed structural and functional analyses of the liver organoids revealed a heterogeneous cellular composition including hepatocyte-, bile duct epithelial-, epithelial-, and stellate-like cells. Structurally, they exhibited bile duct- and hepatic lobule-like formations. Functionally, liver organoids displayed lipid and glycogen accumulation, ICG uptake and release, albumin and urea secretion, as well as metabolic responses to APAP and rifampin. Consequently, our study introduces a high-throughput manufacturing platform for hiPSC-derived organoids, with the potential to generate functional organoids for therapeutic applications and drug screening.