In Vitro Human Liver Model of Nonalcoholic Steatohepatitis by Coculturing Hepatocytes, Endothelial Cells, and Kupffer Cells.
Ceri-Anne E SuurmondSoufian LasliFloor W van den DolderAly UngHan-Jun KimPraveen BandaruKangJu LeeHyun-Jong ChoSamad AhadianNureddin AshammakhiMehmet R DokmeciJunmin LeeStefano PasseriniPublished in: Advanced healthcare materials (2019)
The liver has a complex and unique microenvironment with multiple cell-cell interactions and internal vascular networks. Although nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease with multiple phases, no proper model could fully recapitulate the in vivo microenvironment to understand NAFLD progression. Here, an in vitro human liver model of NAFLD by coculturing human hepatocytes, umbilical vein endothelial cells (HUVECs), and Kupffer cells (KCs) into spheroids is presented. Analysis of indirect cross-talk using conditioned media between steatotic spheroids-composed of hepatocellular carcinoma-derived cells (HepG2) and HUVECs-and mouse KCs reveals that the latter can be activated showing increased cell area, elevated production of reactive oxygen species (ROS), and proinflammatory cytokines. Spheroids incorporating human KCs (HKCs) can also be induced into steatotic stage by supplementing fat. Steatotic spheroids with/without HKCs show different levels of steatotic stages through lipid accumulation and ROS production. Steatotic spheroids made from an immortalized hepatic progenitor cell line (HepaRG) compared to those made from HepG2 cells display similar trends of functionality, but elevated levels of proinflammatory cytokines, and improved reversibility of steatosis. The in vitro human liver system proposed makes strides in developing a model to mimic and monitor the progression of NAFLD.
Keyphrases
- endothelial cells
- induced apoptosis
- reactive oxygen species
- high glucose
- cell cycle arrest
- single cell
- cell death
- stem cells
- cell therapy
- dna damage
- endoplasmic reticulum stress
- adipose tissue
- metabolic syndrome
- type diabetes
- oxidative stress
- insulin resistance
- high fat diet
- vascular endothelial growth factor
- signaling pathway
- diabetic rats
- pluripotent stem cells