Extracellular matrices modulate differentiation of human embryonic stem cell-derived hepatocyte-like cells with spatial hepatic features

Abstract

Abstract Background Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells (HLCs) can provide a valuable in vitro model for disease modelling and drug development. However, it is challenging to generate these cells with functions comparable to hepatocytes in vivo. Extracellular matrices (ECM) play an important role in supporting liver development and hepatocyte functions, but their functions in hepatocyte differentiation and maturation during hPSC differentiation remain unclear. Here, we investigate the effects of two ECM - Matrigel and type I collagen on hepatic differentiation of human embryonic stem cells (hESCs). Methods hESCs-derived HLCs were generated through multistage differentiation in 2D and 3D cultures, incorporating either type I collagen or Matrigel during hepatic specification and maturation. The resulting cells were characterized with various molecular and cellular techniques for their hepatic functionality. Results Our results showed that HLCs cultured with collagen exhibited a significant increase in albumin and alpha 1 anti-trypsin accompanied with reduced AFP compared to HLCs cultured with Matrigel and that they also secreted more urea than Matrigel cells. However, these HLCs exhibited lower CYP3A4 activity and glycogen storage than those cultured with Matrigel. The functional differences in HLCs between collagen and Matrigel cultures closely resembled the hepatocytes of periportal and pericentral zones, respectively. Conclusion Our study demonstrates that Matrigel and collagen have differential effects on the differentiation and functionality of HLCs, which resemble, to an extent, hepatic zonation in the liver lobules. Our finding has an important impact on the generation of hPSC-HLCs for biomedical and medical applications.