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In vitro differentiation effect of CCL4-induced liver injured mice serum on bone marrow-derived mesenchymal stem cells toward hepatocytes like cells

机译:CCL4诱导的肝脏受损小鼠血清对骨髓间充质干细胞对肝细胞的体外分化作用

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Liver dysfunction is a major health problem worldwide. Stem cells therapy has opened up new avenues for researches to treat liver diseases due to their multi lineage differentiation. As mesenchymal stem cells (MSCs) can be differentiated into hepatic lineages in the presence of different exogenous factors, the current study aimed to investigate the impact of carbon tetrachloride (CCl4) induced liver injured mice serum on MSCs differentiation toward hepatocytes in vitro. Male Balb/c mice were treated for liver injury with CCl4 as determined through biochemical tests spectrophotometrically and different growth factors (EGF, HGF) quantification through Sandwich ELISA in both normal and CCl4-induced liver injured mice serum. Mice bone marrow derived-MSCs at second passage were treated with normal and CCl4-induced liver injured mice serum. After 7 days, serum treated MSCs were investigated for hepatocytes like characteristics through RT-PCR. Serum biochemical tests (Bilirubin, ALT and ALP) and sandwich ELISA results of EGF and HGF showed marked increase in CCl4 treated mice serum as compared to normal mice serum(.) Periodic acid Schiff's staining and urea assay kit confirmed high level of glycogen storage and urea production in cells treated with CCl4-induced liver injured mice serum. RT-PCR results of CCl4-induced liver injured mice serum treated cells also showed expression of hepatic markers (Albumin, Cyto-8, Cyto-18, and Cyto-19). This study confirmed that CCl4-induced liver injured serum treatment can differentiate MSCs into hepatocyte-like cells in vitro.
机译:肝功能不全是世界范围内的一个主要健康问题。由于肝脏疾病的多谱系分化,干细胞疗法为治疗肝脏疾病的研究开辟了新的途径。由于间充质干细胞(MSCs)可以在不同外源性因子的作用下分化为肝细胞系,本研究旨在探讨四氯化碳(CCl4)诱导的肝损伤小鼠血清对MSCs体外向肝细胞分化的影响。在正常和CCl4诱导的肝损伤小鼠血清中,用分光光度法测定的生化试验和通过夹心ELISA定量的不同生长因子(EGF、HGF)测定的CCl4治疗雄性Balb/c小鼠的肝损伤。用正常和CCl4诱导的肝损伤小鼠血清处理第二代小鼠骨髓间充质干细胞。7天后,通过RT-PCR检测血清处理的MSC的肝细胞样特征。血清生化试验(胆红素、ALT和ALP)以及EGF和HGF的夹心ELISA结果显示,与正常小鼠血清相比,CCl4处理的小鼠血清显著增加(.)高碘酸希夫染色和尿素分析试剂盒证实,在用CCl4诱导的肝损伤小鼠血清处理的细胞中,糖原储存和尿素生成水平较高。四氯化碳诱导的肝损伤小鼠血清处理细胞的RT-PCR结果也显示肝标记物(白蛋白、细胞因子-8、细胞因子-18和细胞因子-19)的表达。本研究证实,四氯化碳诱导的肝损伤血清治疗可在体外将骨髓间充质干细胞分化为肝样细胞。

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