3D human liver tissue from pluripotent stem cells displays stable phenotype in vitro and supports compromised liver function in vivo

Abstract

Liver disease is an escalating global health issue. While liver transplantation is an effective mode of therapy, patient mortality has increased due to the shortage of donor organs. Developing renewable sources of human liver tissue is therefore attractive. Pluripotent stem cell-derived liver tissue represents a potential alternative to cadaver derived hepatocytes and whole organ transplant. At present, two-dimensional differentiation procedures deliver tissue lacking certain functions and long-term stability. Efforts to overcome these limiting factors have led to the building of three-dimensional (3D) cellular aggregates. Although enabling for the field, their widespread application is limited due to their reliance on variable biological components. Our studies focused on the development of 3D liver tissue under defined conditions. In vitro generated 3D tissues exhibited stable phenotype for over 1 year in culture, providing an attractive resource for long-term in vitro studies. Moreover, 3D derived tissue provided critical liver support in two animal models, including immunocompetent recipients. Therefore, we believe that our study provides stable human tissue to better model liver biology ‘in the dish’, and in the future may permit the support of compromised liver function in humans.Electronic supplementary materialThe online version of this article (10.1007/s00204-018-2280-2) contains supplementary material, which is available to authorized users.

Recapitulating human tissue damage, repair, and fibrosis with human pluripotent stem cell-derived organoids

“…The cells obtained following those protocols possess characteristics of immature, fetal hepatocytes, and fail to maintain the hepatocyte phenotype, limiting their potential for therapeutic or clinical applications . Alternative protocols for the development of human liver tissues relied on matrix or scaffolds for the formation of 3D aggregates with or without inclusion of other cell types . The hPSC‐derived hepatocytes expressed hepatocyte markers including the P450 cytochrome enzymes CYP3A4, CYP3A7, CYP2D6, CYP2C9, and CYP2C19 .…”

“…The hPSC‐derived hepatocytes expressed hepatocyte markers including the P450 cytochrome enzymes CYP3A4, CYP3A7, CYP2D6, CYP2C9, and CYP2C19 . Although hPSC‐derived hepatocytes displayed great potential in transplantation and liver regeneration in mice, the spheroids were not tested in experiments of injury and repair in vitro, and therefore no information is available on their intrinsic mechanisms of repair …”

“…55 Alternative protocols for the development of human liver tissues relied on matrix or scaffolds for the formation of 3D aggregates with or without inclusion of other cell types. [56][57][58][59] The hPSC-derived hepatocytes expressed hepatocyte markers including the P450 cytochrome enzymes CYP3A4, CYP3A7, CYP2D6, CYP2C9, and CYP2C19. 48 Although hPSC-derived hepatocytes displayed great potential in transplantation and liver regeneration in mice, the spheroids were not tested in experiments of injury and repair in vitro, and therefore no information is available on their intrinsic mechanisms of repair.…”

“…48 Although hPSC-derived hepatocytes displayed great potential in transplantation and liver regeneration in mice, the spheroids were not tested in experiments of injury and repair in vitro, and therefore no information is available on their intrinsic mechanisms of repair. 59 Liver fibrosis is a major impairment for hepatocyte-driven liver regeneration, 47 and therefore, it is important to understand the process of liver fibrogenesis. HSCs are the main fibrogenic liver resident cell.…”

Highlight report: role of HNF4α in stem-cell differentiation to hepatocytes

“…Currently, there is a need of in vitro systems with cells that closely resemble mature human cells, such as hepatocytes (Godoy et al 2013(Godoy et al , 2016Ghallab et al 2016;Hewitt et al 2007), kidney epithelial tubular cells (Sjögren et al 2018;Jiang et al 2018;Leist et al 2017), and neuronal cells (Krug et al 2013;Waldmann et al 2014;Rempel et al 2015). Human hepatic differentiation protocols have been developed and optimized (Cameron et al 2015;Rashidi et al 2018;Sullivan et al 2010), but the problem remains that stem-cell-derived hepatocyte-like cells still show major differences compared to primary human hepatocytes. One common problem is too low expression of hepatocyte-specific genes; a further challenge is expression of 'unwanted genes' that are not observed in primary human hepatocytes (Sachinidis et al 2019;Godoy et al 2015).…”