Multilineage communication regulates human liver bud development from pluripotency.

Conventional two-dimensional differentiation from pluripotency fails to recapitulate cell interactions occurring during organogenesis. Three-dimensional organoids generate complex organ-like tissues; however, it is unclear how heterotypic interactions affect lineage identity. Here we use single-cell RNA sequencing to reconstruct hepatocyte-like lineage progression from pluripotency in two-dimensional culture. We then derive three-dimensional liver bud organoids by reconstituting hepatic, stromal, and endothelial interactions, and deconstruct heterogeneity during liver bud development. We find that liver bud hepatoblasts diverge from the two-dimensional lineage, and express epithelial migration signatures characteristic of organ budding. We benchmark three-dimensional liver buds against fetal and adult human liver single-cell RNA sequencing data, and find a striking correspondence between the three-dimensional liver bud and fetal liver cells. We use a receptor-ligand pairing analysis and a high-throughput inhibitor assay to interrogate signalling in liver buds, and show that vascular endothelial growth factor (VEGF) crosstalk potentiates endothelial network formation and hepatoblast differentiation. Our molecular dissection reveals interlineage communication regulating organoid development, and illuminates previously inaccessible aspects of human liver development.

Increased mitochondrial superoxide dismutase expression and lowered production of reactive oxygen species during rotavirus infection.

Rotaviruses are responsible for severe diarrhea in infants and substantial economic losses in animal husbandry worldwide. We investigated the oxidant/antioxidant status in rotavirus-infected human colon adenocarcinoma (Caco-2) cell line. Our results show that within the initial 48 h of infection the expression of the mitochondrial superoxide dismutase (MnSOD) is significantly increased, which correlates with a decrease in reactive oxygen species production, and with a lack of cellular glutathione depletion. During this period the mitochondria display a hyperpolarization of the inner membrane, which leads to an increased mitochondrial membrane potential. No increase in apoptosis was detected in the infected cultures. In contrast to many viral infections which cause redox imbalance in host cells, the described virus-host interaction suggests that rotavirus infection does not lead to an induction of oxidative stress, possibly to prolong cell survival and to allow for accumulation of viral particles before cell destruction and virus release.