Biochemical and Ultrastructural Studies on the Effect of Human Mesenchymal Stem Cells on Hepatoma Cell Line (HuH7)

Abstract

This study was designed to isolate mesenchymal stem cells from human umbilical cord for treatment of hepatocellular carcinoma cell line (HuH7) and to evaluate the efficiency of using two media (conditioned and co-culture media) and their effect on morphological, ultrastructure,biochemical and genetic parameters . The media affected greatly morphology and cell viability .Electron microscope showed great effect on HuH7 such as high vacuolation, loss of RER and ribosomes, decrease in number of intact mitochondria and increase of lipid droplets,apoptotic bodies and cell fragments .Biochemical parameters measured were Alfa fetoprotein as tumor marker, albumin, GOT, GPT, Alkaline phosphatase andGT in HuH7cultured in normal media and HuH7 cultured in conditioned and co-culture media at different time intervals (0, 48hrs, 72hrs and 96hrs). Data showed significant difference in liver enzyme (GOT), alfa fetoprotein and non-significant difference in albumin, ALP and GT between cells in normal media and those grown in conditioned or in co-culture media . Gene expression changes for Survivin, PCNA, β-Catenin, Telomerase and VEGF in HuH7 was analysed in this study and demonstrated a significant down regulation in Survivin (p<0.001), PCNA (p<0.001), VEGF (p<0.001) and ßCatenin (p=0.022) but not significant in Telomerase (p=0.617).