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

Abstract

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.

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Acknowledgements

The authors would like to thank Abdul Wali Khan University Mardan, Pakistan for providing financial support and research facilities for this research work.

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No funding received for the current study.

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Correspondence to Abdul Nasir.

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The authors declare that they have no conflicts of interest.

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All the manipulations with animals were carried out in accordance to the informed consent protocol approved by the Ethics Committee of the Abdul Wali Khan University Mardan. This article does not contain any studies with human participants performed by any of the authors.

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Shams, S., Imran, N., Afridi, S.G. et al. In vitro differentiation effect of CCL4-induced liver injured mice serum on bone marrow-derived mesenchymal stem cells toward hepatocytes like cells. Cell Tissue Bank (2020). https://doi.org/10.1007/s10561-020-09878-5

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Keywords

  • Liver
  • Stem cells
  • Ccl4-induced injured serum
  • Growth factors
  • Hepatocytes differentiation