ABSTRACT

          Background and Objective: Multipotent placental amniotic membrane mesenchymal stem cells (MSCs) are capable of differentiating into specialized tissues under different conditions. The aim of this study was to induce differentiation of placental amniotic membrane MSCs from NMRI mouse into hepatocytes using liver extract.

         Methods: Placental amniotic membrane MSCs from a 14-day pregnant female mouse was used in this study. The cells were incubated with trypsin solution, followed by pipetting. The resulting suspension was cultured in 12-well plates. After confirming their mesenchymal nature, differentiation of the aforementioned cells was induced via exposure to 6, 18, 30 and 60 μg/ml of liver extract. On the 16^th day of treatment, immunocytochemical reaction for albumin and periodic acid-Schiff (PAS) test were performed for detection of hepatocyte-like cells.

          Results: Change was observed in the shape of differentiating cells from spindle-like shape to polygonal shape. The immunocytochemical reaction of the differentiated cells was positive. PAS staining also confirmed the accumulation of glycogen particles in the aforementioned cells. Concentration of 6 μg/ml liver extract was found as the effective dose for induction of differentiation.

           Conclusion: The findings of this study show that the placental amniotic membrane-derived MSCs of mouse can differentiate in vitro from spindle-like cells to polygonal hepatocyte-like cells with large nuclei and under the influence of the liver.

Keywords: Placental Amniotic Membrane Mesenchymal Stem Cells, Hepatocyte, In Vitro.

ABSTRACT
          Background and objectives: Tricyclazole (TCZ) is a member of triazole fungicides, which might cause damage in living systems. This study was carried out to examine effects of TCZ on liver tissues and level of liver enzymes.
            Methods: Forty mice were randomly divided into four groups including control, sham and two experimental groups. Experimental groups 1 and 2 received 5 mg/Kg and 15 mg/Kg intraperitoneal injection of TCZ for two weeks, respectively. The sham group received sterile water but the control group received no injection. The animals were sacrificed 24 h after the last injection, and microscopic slides were prepared for cell counting and evaluation of tissue damage. Levels of liver enzymes were measured using commercial kits. Data was analyzed in SPSS (version 20) using one-way ANOVA.
          Results: The injection of TCZ caused a significant increase in the number of hepatocytes and a significant decrease in the number of Kupffer cells compared to control group (P<0.001). In the experimental group, the level of alanine aminotransferase and aspartate aminotransferase increased, but the level of alkaline phosphatase decreased significantly compared to control group (P<0.001). We also detected several forms of tissue damage including necrosis and degeneration of hepatocytes, hyperplasia, and penetration of inflammatory cells and expansion of sinusoids.
          Conclusion: Our results indicate that the intraperitoneal injection of TCZ in mice can cause irreparable hepatic damage in a dose-dependent manner.
          Keywords: Tricyclazole, hepatocytes, Alanine, Aspartate aminotransferase.