Leveraging complex interactions between signaling pathways involved in liver development to robustly improve the maturity and yield of pluripotent stem cell-derived hepatocytes

Abstract

SUMMARYPluripotent stem cell (PSC)-derived hepatocyte-like cells (HLC) have shown great potential as an alternative to primary human hepatocytes (PHH) for in vitro modeling. Several differentiation protocols have been described to direct PSC towards the hepatic fate, although the resulting HLC have shown more a fetal than adult phenotype. Here, by leveraging recent knowledge of the signaling pathways involved in liver development, we describe a robust, scalable protocol that allows to consistently generate high-quality HLC from both ESC and iPSC. Such HLC are comparable to adult PHH in terms of key mature liver functions and proved suitable to assess drug hepatotoxicity, as a proof of concept of their potential as a physiologically representative alternative for in vitro modeling.

Autologous humanized mouse models of iPSC-derived tumors enable characterization and modulation of cancer-immune cell interactions

“…To generate alternative tumor models that can be used in a humanized setting, we set out to transform iPSC-derived cells from healthy donors. First, iPSCs were differentiated into hepatocytes by using a previously described protocol (schematized in Figure 3 A) ( Raggi et al., 2020 ). We initiated the transformation process using SV40ER at different time points in the cell differentiation process, which led to the formation of growing colonies.…”

“…Differentiation of human iPSCs into hepatocyte-like cells (HLC) was achieved according to our 30-day long, previously described protocol mimicking liver development ( Raggi et al., 2020 ). In brief, iPSCs were differentiated on laminin-coated plates through consecutive stages (primitive streak, mesendoderm, definitive endoderm, posterior foregut, hepatoblasts and, eventually, HLC).…”

Autologous humanized mouse models of iPSC-derived tumors allow for the evaluation and modulation of cancer-immune cell interactions

“…First, iPSC were differentiated into hepatocytes using a previously described protocol (schematized in Fig. 3A) (24). We initiated the transformation process using SV40ER at different timepoints in the cell differentiation process which led to the formation of growing colonies.…”

“…Differentiation of human iPSCs into hepatocyte-like cells (HLC) was achieved according to our 30day long, previously described protocol mimicking liver development (24). In brief, iPSCs were differentiated on laminin-coated plates through consecutive stages (primitive streak, mesendoderm, definitive endoderm, posterior foregut, hepatoblasts and, eventually, HLC).…”