Development of Simple Protocol for Generation of Functionally Active Hepatocyte-like Cells from Human Adipose Tissue-derived Stem Cells
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Abstract:
Background and Aims: Human adipose tissue-derived stem cells (hASCs) are considered as an attractive source of regenerative stem cells, mainly because of their higher proliferation rate, more accessibility and hepatocyte like properties as compared to mesenchymal stem cells isolated from other tissues. Numerous studies have described the beneficial use of adipose tissue-derived stem cells for generating hepatocyte-like cells. However, due to the lack of appropriate culture conditions, most of the produced cells exhibit poor functionality. The aim of the present study was to establish a new protocol for the efficient hepatic differentiation of hASCs. Materials and Methods: hASCs were cultured in hepatic differentiation medium containing fibroblast growth factor 4, hepatocyte growth factor, dexamethasone and oncostatin M using a three-step protocol up to 21 days. Then, the functionality of the treated cells was evaluated by analyzing specific hepatocyte genes and biochemical markers at various time points. Results: A significant upregulation in albumin, alpha-fetoprotein, cytokeratin 18 and hepatocyte nuclear factor-4α expressions was observed in differentiated cells relative to day 1 of differentiation protocol. Moreover, the finding of glycogen deposits increased urea production and positive immunofluorescence staining for albumin and alpha-fetoprotein in hepatocyte-like cells suggesting that most of the cells differentiate into hepatocyte-like cells. Conclusions: Our report has provided a simple protocol for differentiation of hASCs into more functional hepatocyte-like cells.
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Journal title
volume 6 issue 1
pages 51- 62
publication date 2019-02
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