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Localizing, identifying and quantifying lipid droplets are important features to be assessed in the research of mechanisms involved in diseases such as steatosis, obesity, diabetes, myopathies and arteriosclerosis. Lipid-droplet staining with Oil Red O is widely used combined to either light field, conventional fluorescence microscopy and phase contrast microscopy. Here, for the first time, we report an easy, fast and precise protocol for the quantitative evaluation of lipid droplets staining with Oil Red O in HepG2 cells in vitro using confocal laser scanning microscopy associated with maximum intensity projection technique and counting point method. Our methodology was compared to a previous described protocol to measure lipid droplets, based on phase contrast microscopy and binarization. Our protocol substantially enhanced the quality of lipid droplets images compared to phase contrast microscopy by different reasons, such as: 1-Use of maximum intensity projection technique; 2-Out-of-focus light absence; 3-Increased contrast; 4-Enhanced lipid droplet definition. Thus, the use of this protocol increases the sensitivity of lipid droplets quantification, showing morphological results that can be underestimated using other approaches. [ABSTRACT FROM AUTHOR]

The incidence of hepatocellular carcinoma (HCC) keeps rising year by year, and became the second leading cause of cancer-related death. Some studies have found that liraglutide, a GLP-1 analog, may decrease the tumor cells proliferation. Due to this, the aim of this work is to investigate the antiproliferative potential of exenatide, another GLP-1 analog. Cell proliferation was assessed by direct count with Trypan blue dye exclusion. Flow cytometry was used to determinate autophagy and nuclear staining. Morphometric analysis was used to verify senescence and apoptosis. The mechanism that induced cell growth inhibition was analyzed by Western Blot. Treatment with exenatide significantly decreases cell proliferation and increases autophagy, both in relation to control and liraglutide. In addition, mTOR inhibition was greater in cells treated with exenatide. In relation to chronic treatment, exenatide does not allow cellular regrowth by preventing some resistance mechanism that the cells can acquire. These results suggest that exenatide has a potent anti-proliferative activity via mTOR modulation and, among the GLP-1 analogs tested, could be in the future an alternative for HCC treatment., (Copyright © 2019 Krause et al.)