Transient culture of adult human hepatocytes as organoids improves engineered liver engraftment.

Bioengineered livers could serve as a bridge or alternative to whole organ transplantation for patients suffering from liver disease. However, identifying cellular configurations that promote adult human hepatocyte survival after tissue transplantation remains a critical hurdle. We hypothesized that transient culture of adult hepatocytes in 3D as organoids would enhance engraftment of these cells in bioengineered tissues in vivo. Here, we established an organoid system that supports culture of adult human hepatocytes from eight different human adult donors aged 19-49. These organoids support retention of human hepatocyte morphology found in the mature liver, express an adult liver transcriptome, and exhibit mature functions such as inducible cytochrome activity. Importantly, adult human hepatocyte organoids (AHHO) improved engraftment of hepatocytes up to 25-fold in vivo in engineered tissues compared to previous hepatocyte models. Thus, 'priming' human cells in organoids could have a major impact on translational bioengineered tissue applications.