Longitudinal scRNA-seq analysis of human liver bud organoid (HLBO) with iPS-derived liver sinusoidal endothelial progenitor (iLSEP)

We directly differentiated human pluripotent stem cells into CD32b+ putative liver sinusoidal progenitors (iLSEP) by dictating developmental pathways. By devising an inverted multilayered air-liquid interface (IMALI) culture, hepatic endoderm, septum mesenchyme, arterial and sinusoidal quadruple progenitors self-organized to generate vascularized liver bud organoid. To uncover cellular characteristics and diversity during organoid development at transcriptomic level, time-series single cell RNA-seq (scRNA-seq) data were acquired. At days 3, 7, and 11 after self-assembly culture, three HLBOs at each time point were subjected to scRNA-seq library preparation.