Single-Cell Transcriptomics of Acetaminophen-Induced Responses in Human 2D and 3D Liver Microtissues

This study aimed to investigate how liver cell culture architecture influences acetaminophen (APAP)-induced transcriptional responses at single-cell resolution. We used human liver microtissues composed of primary human hepatocytes (PHHs), Kupffer cells (KCs), and hepatic sinusoidal endothelial cells (HSECs), cultured as 2D monolayers or 3D spheroids. Microtissues were exposed to vehicle, low-dose (350?µM), or high-dose (2687?µM) APAP for 24?hours. Single-cell suspensions were generated, and RNA libraries were prepared using the 10x Genomics Chromium platform, followed by Illumina sequencing. The resulting data were analyzed using Cell Ranger and Seurat to identify differentially expressed genes, assess pathway enrichment, and characterize cell-type–specific transcriptional responses to APAP exposure.