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Differentiation of hepatocytes from induced pluripotent stem cells derived from human hair follicle mesenchymal stem cells

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Abstract

Due to the limitations of organ donors and immune rejection in severe liver diseases, stem cell-based therapy presents a promising application for tissue repair and regeneration. As a novel cell source, mesenchymal stem cells separated from human hair follicles (HF-MSCs) are convenient to obtain and have no age limit. To date, the differentiation of HF-MSCs into hepatocytes has not been reported. In this study, we explored whether HF-MSCs and HF-MSC-derived-induced pluripotent stem cells (HF-iPS) could differentiate into hepatocytes in vitro. Flow cytometry, Oil Red O stain and Alizarin Red stain were used to identify the characteristics of HF-MSCs. The expression of liver-specific gene was detected by immunofluorescence and Quantitative Polymerase Chain Reaction. Periodic Acid-Schiff stain, Indocyanine Green stain and Low-Density Lipoprotein stain were performed to evaluate the functions of induced hepatocyte-like cells (HLCs). HF-MSCs were unable to differentiate into HLCs using previously reported procedures for MSCs from other tissues. However, HF-iPS efficiently induced the generation of HLCs that expressed hepatocyte markers and drug metabolism-related genes. HF-iPS can be used as novel and alternative cellular tools for inducing hepatocytes in vitro, simultaneously benefiting from utilizing HF-MSCs as a noninvasive and convenient cell source for reprogramming.

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Acknowledgments

We thank LetPub for its linguistic assistance during the preparation of this manuscript.

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Authors and Affiliations

  1. The Key Laboratory of Pathobiology, Ministry of Education, College of Basic Medical Sciences, Jilin University, Changchun, 130021, China

    Xu Shi, Shuang Lv, Xia He, Xiaomei Liu, Meiyu Sun, Meiying Li, Guangfan Chi & Yulin Li

  2. Genetic Diagnosis Center, Central Laboratory, The First Hospital of Jilin University, Changchun, 130032, China

    Xu Shi

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  1. Xu Shi
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  2. Shuang Lv
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Correspondence to Yulin Li.

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Figure S1

Identification of HF-MSCs. a–c Morphology characterization. A typical fibroblast-like cell shape was observed in the undifferentiated HF-MSCs. Adipogenic differentiation was detected by Oil Red O staining. The tiny red dots represent intracellular lipid droplets. Osteogenic differentiation was detected by Alizarin Red staining. A red color indicates the formation of calcium nodules. Scale bars  20 μm. d–j Flow cytometric analyses of cell surface markers on HF-MSCs. A tiotal 2 × 105 HF-MSCs were incubated with primary antibodies against different biomarkers. Controls were incubated with secondary antibody only. % indicates the fraction of cells that stained positive. (GIF 124 kb)

High resolution image (TIF 4284 kb)

Figure S2

Immunofluorescence analysis of DE markers in HF-MSC-derived iPS (HF-iPS). a–i Undifferentiated HF-iPS showed negative staining for DE markers—SOX17, GATA4 and FoxA2. Scale bars 50 μm. (GIF 139 kb)

High resolution image (TIF 4416 kb)

Figure S3

Identification of hepatic functions in HF-MSC-derived iPS (HF-iPS). Transport and metabolic functions, as well as glycogen storage were detected in undifferentiated HF-iPS cells. a–c The detection of LDL uptake in undifferentiated HF-iPS. Nuclei were stained with DAPI (blue). Scale bars 50 μm. d, e PAS staining and ICG uptake analysis in HF-iPS. Scale bars 50 μm. (GIF 33 kb)

High resolution image (TIF 1192 kb)

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Shi, X., Lv, S., He, X. et al. Differentiation of hepatocytes from induced pluripotent stem cells derived from human hair follicle mesenchymal stem cells. Cell Tissue Res 366, 89–99 (2016). https://doi.org/10.1007/s00441-016-2399-5

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  • DOI: https://doi.org/10.1007/s00441-016-2399-5

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