Primary murine hepatocytes exposed to fatty acids analyzed by Raman and infrared microscopy

Aim of this work is to present the phenotype and biochemical characteristics of primary freshly isolated murine hepatocytes exposed to oleic and palmitic acids, the most abundant fatty acids (FAs) in human diet and serum. Excess of FAs in the diet is one of the factors that contributes to the non-alcoholic fatty liver disease (NAFLD). The hallmark of NAFLD is the overaccumulation of lipids in hepatic tissue and formation of lipid droplets (LDs). With time NAFLD can progress into more advanced stages leading to liver injury. Hepatocytes as the most abundant cells in the liver are strongly involved in the process of LDs formation, therefore are widely used in research on molecular mechanism of NAFLD development and progression. Primary hepatocytes are rarely used in investigations, due to multistep isolation process and loss of functionality under culture conditions. However, they offer substantial advantages, including preserved phenotype and functions. In order to investigate LDs formation and their content, the Raman and FTIR imaging were employed, as they provide spatial and vibrational information on studied cells in label-free manner.