In vitro differentiated hepatic oval-like cells enhance hepatic regeneration in CCl4-induced hepatic injury.
Hepatic oval cells are likely to be activated during advanced stage of liver fibrosis to reconstruct damaged hepatic tissue. However, their scarcity, difficulties in isolation, and in vitro expansion hampered their transplantation in fibrotic liver. This study was aimed to investigate the repair potential of in vitro differentiated hepatic oval-like cells in CCl4-induced liver fibrosis. BMSCs and oval cells were isolated and characterized from C57BL/6 GFP+mice. BMSCs were differentiated into oval cells by preconditioning with HGF, EGF, SCF, and LIF and analyzed for the oval cells-specific genes. Efficiency of oval cells to reduce hepatocyte injury was studied by determining cell viability, release of LDH, and biochemical tests in a co-culture system. Further, in vivo repair potential of differentiated oval cells was determined in CCl4-induced fibrotic model by gene expression analysis, biochemical tests, mason trichrome, and Sirius red staining. Differentiated oval cells expressed hepatic oval cells-specific markers AFP, ALB, CK8, CK18, CK19. These differentiated cells when co-cultured with injured hepatocytes showed significant hepato-protection as measured by reduction in apoptosis, LDH release, and improvement in liver functions. Transplantation of differentiated oval cells like cells in fibrotic livers exhibited enhanced homing, reduced liver fibrosis, and improved liver functions by augmenting hepatic microenvironment by improved liver functions. This preconditioning strategy to differentiate BMSCs into oval cell leads to improved survival and homing of transplanted cells. In addition, reduction in fibrosis and functional improvement in mice with CCl4-induced liver fibrosis was achieved.
National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.|Institute of Molecular Biology and Biotechnology, University of Lahore, Lahore, Pakistan.
Sana Javaid
SJ
Awan
National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
Maria Tayyab
MT
Baig
National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
Faiza
F
Yaqub
School of Biological Sciences, University of the Punjab, Lahore, Pakistan.
Asima
A
Tayyeb
National Center of Excellence in Molecular Biology, University of the Punjab, Lahore, Pakistan.
Gibran
G
Ali
D000818:Animals; D002251:Carbon Tetrachloride; D002454:Cell Differentiation; D019070:Cell Lineage; D002470:Cell Survival; D060833:Cellular Microenvironment; D018920:Coculture Techniques; D022781:Hepatocytes / Q000473:pathology*; D008099:Liver / Q000293:injuries* / Q000473:pathology*; D008103:Liver Cirrhosis / Q000473:pathology / Q000628:therapy; D008115:Liver Regeneration; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology; D008810:Mice, Inbred C57BL
D002251:Carbon Tetrachloride
cell death; cell differentiation; cellular imaging; fluorescence techniques; growth factors; hepatocytes