Urine-Derived Stem Cells Repair Liver Function in Nude Mice with Chronic Liver Injury
Hu Chaoqun1,2, Fang Shuyu1, Gong Mengjia1, He Yun1, Bi Yang1*
In the present study, we try to investigate the biological characteristics of urine-derived stem cells (USCs) and its potentiality on the treatment of chronic liver injury. USCs were isolated and cultured, cell morphology was observered under light microscopy. Flow cytology was used to detect the stem cell surface markers. Alkaline phosphatase (ALP) staining, alizarin red staining, oil red O staining, indocyanine green (ICG) uptake test and PAS (periodic acid-Schiff) staining were performed to evaluate osteogenic, adipogenic, and hepatic differentiation of USCs. The nude mice model of chronic liver injury was established by intraperitoneal injection of carbon tetrachloride (CCL4), and then USCs were transplanted into mice model by tail vein injection. Liver index, ALT, AST in serum, HE and Masson staining were performed to determine the therapeutic effect. The results showed that USCs were ricelike adherent growth cells expressing various mesenchymal stem cell markers, including CD24, CD29, CD73, CD90 and CD105, the periodic surface marker CD146, and not expressing hematopoietic cell surface markers CD31, CD34, CD45. With osteogenic and adipogenic induction, some of treated USCs were positive for ALP staining, alizarin red staining and oil red O staining. With hepatic induction alone, almost no USCs has the function of ICG uptake and no positive PAS staining cells, while 10% of USCs were positive for ICG uptake and PAS staining when induced and co-cultured with hepatic progenitor cells. The liver index of the transplanted group was lower than that of the model group, however, there was no statistically significance of ALT and AST between two groups. H.E and Masson staining showed that the degeneration of hepatocytes and the fibrosis in the transplantation group was improved compared with that in the model group. Therefore, we successfully isolated and cultured USCs which has the abilities of good proliferation and multiple differentiation. Transplantion of USCs could partly repair liver injury and pathology of chronic liver injury mice model.
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