Stem Cell Regenerative Medicine Lab, Department of Biochemistry, Abdul Wali Khan University Mardan, Mardan, 23200, Khyber Pakhtunkhwa, Pakistan.
Department of Biological Sciences, National University of Medical Sciences, Rawalpindi, 46000, Pakistan.
Cell Tissue Bank. 2021 Jun;22(2):297-303. doi: 10.1007/s10561-020-09878-5. Epub 2020 Nov 10.
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 (CCl) induced liver injured mice serum on MSCs differentiation toward hepatocytes in vitro. Male Balb/c mice were treated for liver injury with CCl as determined through biochemical tests spectrophotometrically and different growth factors (EGF, HGF) quantification through Sandwich ELISA in both normal and CCl-induced liver injured mice serum. Mice bone marrow derived-MSCs at second passage were treated with normal and CCl-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 CCl 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 CCl-induced liver injured mice serum. RT-PCR results of CCl-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 CCl-induced liver injured serum treatment can differentiate MSCs into hepatocyte-like cells in vitro.
肝功能障碍是全球范围内的一个主要健康问题。由于其多谱系分化，干细胞疗法为治疗肝脏疾病的研究开辟了新途径。由于间充质干细胞 (MSCs) 在存在不同外源性因素的情况下可以分化为肝谱系，因此本研究旨在探讨四氯化碳 (CCl) 诱导的肝损伤小鼠血清对 MSCs 向体外肝细胞分化的影响。通过比色分光光度法测定雄性 Balb/c 小鼠的生化试验和正常及 CCl 诱导的肝损伤小鼠血清中的不同生长因子 (EGF、HGF) 的定量，用 CCl 处理以诱导肝损伤。第 2 代骨髓来源的-MSCs 用正常和 CCl 诱导的肝损伤小鼠血清处理。7 天后，通过 RT-PCR 检测血清处理的 MSCs 是否具有肝细胞样特征。血清生化试验 (胆红素、ALT 和 ALP) 和 EGF 和 HGF 的夹心 ELISA 结果表明，与正常小鼠血清相比，CCl 处理的小鼠血清中的这些因子明显增加。过碘酸希夫染色和尿素试剂盒证实，用 CCl 诱导的肝损伤小鼠血清处理的细胞中储存了大量的糖原并产生了尿素。用 CCl 诱导的肝损伤小鼠血清处理的细胞的 RT-PCR 结果还显示出肝脏标志物 (白蛋白、细胞色素 8、细胞色素 18 和细胞色素 19) 的表达。本研究证实，CCl 诱导的肝损伤血清处理可在体外将 MSCs 分化为肝细胞样细胞。
