Hepatotoxicity evaluation of co-exposure of microplastics and TBBPA by 3D biofabricated multicellular organoids

We combined human induced pluripotent stem cells (hiPSCs) and 3D printing to mass-produced liver Disse space-like organoids in microspheres hydrogels (DOs). After 5 days of co-culture, the cells within the microspheres were aggregated and developed into liver organoids, composed of iPSC-derived hepatocytes, endothelial cells, and hepatic stellate cells (LX2). These organoids mirrored the hepatotoxicity of polystyrene microplastics (PS-MPs), TBBPA, and complexes MPTA with healthy and patient-derived organoids. Overall design: Healthy person hiPSCs (N_iPSCs) and alcoholic liver disease patient hiPSCs (AC19-iPSCs) were differentiated into hepatic endoderm and endothelial cells. With hepatic endoderm, endothelial cells, LX-2 and alginate / laminin511, we fabricated healthy and patient-derived liver Disse space-like organoids: N_DOs and AC19_DOs. N_DOs were incubated with 60 ng/mL of polystyrene microplastics (MPs)(named N_MPs), 100 nM of TBBPA (named N_TBBPA), MPTA at 60 ng/mL/100 nM (N_MPTA), and 0.01% DMSO (N_Ctrl) for 3 days. The AC19_DOs were exposed to MPTA at 60 ng/mL/100 nM (AC19_MPTA) and 0.01% DMSO (AC19_Ctrl). The media were refreshed every day. To reveal the features of N_DOs and AC19_DOs, we compared them with the corresponding pre-printed cell mixtures (N_BPs and AC19_BPs) and primary human hepatocytes (PHH).