Hepatocyte-like cells derived from induced pluripotent stem cells inhibit liver fibrosis in rats
10.3969/j.issn.2095-4344.1607
- VernacularTitle:诱导多功能干细胞来源肝样细胞体内抗肝纤维化
- Author:
Gang CHENG
1
;
Denggao HUANG
;
Ying LIANG
Author Information
1. 中南大学湘雅医学院附属海口医院肿瘤放疗科
- Keywords:
Liver Cirrhosis;
Induced Pluripotent Stem Cells;
Hepatocytes;
Tissue Engineering
- From:
Chinese Journal of Tissue Engineering Research
2019;23(12):1384-1389
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Induced pluripotent stem cells have similar self-renewal, proliferation and differentiation abilities to embryonic stem cells. They have no source limitations, no ethical problems, and no current problems of cell xenogenesis, which are expected to be the source of cells for the treatment of liver diseases. OBJECTIVE: To observe the effect of induced pluripotent stem cell-derived hepatocyte-like cells on liver fibrosis. METHODS: The three-step method in vitro was used to induce the differentiation of induced pluripotent stem cells into hepatocyte-like cells. Glycogen staining, immunohistochemistry and low-density lipoprotein uptake assay were used to detect the ability of induced cells to synthesize glycogen, alpha-fetoprotein, albumin, CK18 protein and low-density lipoprotein uptake. Forty-five Sprague-Dawley rats (provided by the Experimental Animal Center, the Affiliated Haikou Hospital, Xiangya School of Medicine, Central South University) were randomized into three groups: normal control group, model group and cell transplantation group (n=15 per group). The rats in the latter two groups were intraperitoneally injected with carbon tetrachloride to establish liver fibrosis models. Cell transplantation group was given intravenous injectin of hepatocyte-like cells (induced for 21 days) , 0.5 mL, 2×109/L, at 3 days after modeling. Four weeks after cell transplantation, venous blood and liver tissue samples were taken to analyze the changes of liver function, liver fibrosis index and liver pathology. RESULTS AND CONCLUSION: (1) After 21 days of induction, human induced pluripotent stem cell clonal clusters became loose, mainly round or polygonal in shape, and presented with a dense paving stone-like arrangement. A large amount of pink glycogens could be seen in the cytoplasm, indicating that induced pluripotent stem cells have the ability to synthesize glycogen. Low-density lipoprotein uptake test showed that induced pluripotent stem cells had the ability to uptake low-density lipoprotein. Immunohistochemistry detection showed that the cells were positive for alpha fetoprotein, albumin and CK18. (2) At 4 weeks after cell transplantation, the level of albumin in the model group was significantly lower than that in the normal control group (P < 0.05) , while the levels of direct bilirubin, indirect bilirubin, aspartate aminotransferase, alanine aminotransferase, type IV collagen, serum hyaluronidase, serum type III procollagen in the model group were significantly higher than those in the normal control group (P < 0.05). Compared with the model group, the level of albumin in the cell transplantation group was significantly increased (P < 0.05) , while the levels of direct bilirubin, indirect bilirubin, aspartate aminotransferase, alanine aminotransferase, type IV collagen, serum hyaluronidase, serum type III procollagen in the cell transplantation group were significantly decreased (P < 0.05). (3) Four weeks after cell transplantation, inflammatory cell infiltration, hepatocyte degeneration and necrosis in the cell transplantation group were improved to different extents. Therefore, hepatocyte-like cells derived from induced pluripotent stem cells could significantly improve liver fibrosis in rats.
