Use of three‐dimensional spheroids of hepatocyte‐derived reporter cells to study the effects of intracellular fat accumulation and subsequent cytokine exposure

摘要: Non-alcoholic fatty liver disease (NAFLD) is a family of diseases associated with obesity. Initial stage NAFLD characterized by liver, referred to as steatosis, which progresses in some individuals non-alcoholic steatohepatitis (NASH) and failure. In order study treat the many such NAFLD, an improved vitro cellular model needed. Several studies past have attempted elucidate these mechanisms using primary hepatocytes or relevant hepatoma cell lines two-dimensional (2D) monolayer cultures. These 2D planar culture systems, unfortunately, do not represent complex architecture hepatic tissue vivo. Therefore, we engineered elastin-like polypeptide (ELP)-polyethyleneimine (PEI) copolymer shown that ELP-PEI coated surfaces influenced H35 rat morphology create 3D spheroids. Our reporter recapitulates features human disease, including acid uptake, intracellular triglyceride accumulation, decreased proliferation, liver-specific function, increased reactive oxygen species accumulation. Finally, demonstrate utility cells for studying transcriptional regulation, compared dynamics nuclear factor κB (NFκB) response its classical inducer (tumor necrosis factor-α, TNF-α) under lean conditions both configurations. We found that, spheroids, linoleic treatment activated NFκB at earlier time points during development but suppressed TNF-mediated activation later points. therefore provide good starting point evaluate relationships observed culture.