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In vitro generation of myofibroblasts-like cells from liver epithelial progenitor cells of rhesus monkey (Macaca mulatta)

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Abstract

The origin of the myofibroblast, the primary effector cell of liver fibrosis, is still elusive. Here, we report that fluorescence-activated cell sorting purified E-cad + rhesus monkey liver epithelial progenitor cells (mLEPCs) may serve as a potential source for liver myofibroblasts. Adult mLEPCs colonies were cultured in medium containing 2 ng/ml transforming growth factor β (TGF-β) and 10% fetal bovine serum (FBS) to induce differentiation. Phenotypic changes of cells were analyzed by morphological observation, immunostaining, and reverse transcription-polymerase chain reaction (RT-PCR). After cultured with TGF-β and FBS, some cells in adult mLEPCs colonies converted to fibroblasts-like cells. Immunostaining showed that fibroblasts-like cells had acquired the expression of mesenchymal cell marker vimentin but lost the expression of epithelial cell marker CK8. Fibroblasts-like cells were maintained in culture for up to 40 passages. RT-PCR analysis revealed that fibroblasts-like cells had acquired the expression of mesenchymal genes (snail, PAI-1, and collagen I) and lost the expression of epithelial specific genes (E-cad, ZO-1, CK18, and occludin). In addition, more than 60% of fibroblasts-like cells expressed myofibroblastic-related proteins such as αSMA, vimentin, and N-cad, which were not presented in mLEPCs. Furthermore, increased cell motility was also detected in these fibroblasts-like cells by time-lapse video observation. Our results demonstrate that hepatic epithelial progenitor cells, mLEPCs, transform to myofibroblast-like cells via epithelial-mesenchymal transition. This finding will facilitate understanding of the origin of myofibroblasts in liver fibrosis.

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Acknowledgements

We wish to acknowledge Mr. Li Jian for assistance in confocal microscopic analysis. This work was supported by research grants from Zhejiang Provincial Natural Science Foundation of China Y2110911, National 863 High Technology Research and Development Program of China 2005AA219010, 2006AA02A101, and 2006AA02A116, Major State Basic Development Program 2006CB701505 and 2007CB947701, The Chinese Academy of Sciences KSCX1-YWR-47 and KSCX1-05-02, R&D Infrastructure and Facility Development Program of Yunnan Province 2006PT08-2, Social Science and Technology Development Program of Yunnan Province 2007GH, National S&T Major Project 2009ZX09501- 028, National Key Technology R&D Program 2007BAI33B00, and Chinese National Science Foundation 30700159 and 30700425.

Conflicts of Interest

None declared.

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Authors and Affiliations

  1. College of Life Science of Shaoxing University, 900# Chennan Dadao, Shaoxing, Zhejiang, 312000, China

    Shaohui Ji & Lifang Jin

  2. School of Life Science of Zhejiang Sci-Tech University, Hangzhou, Zhejiang, China

    Jianhong Shu

  3. Kunming Primate Research Center, and Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan, China

    Shaohui Ji, Xihong Wang, Wei Si, Xiangyu Guo, Bo Zhao, Weizhi Ji & Lifang Jin

  4. Department of Pathology, Shaoxing People’s Hospital and the First Affiliated Hospital of Shaoxing University, Shaoxing, Zhejiang, China

    Aijing Sun

  5. Graduate School, Chinese Academy of Sciences, Beijing, China

    Xihong Wang & Bo Zhao

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  1. Shaohui Ji
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Correspondence to Lifang Jin.

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Editor: T. Okamoto

Shaohui Ji and Xihong Wang contributed equally to this work.

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Supplemental Figure

Cell movement of the mLEPCs derived fibroblasts-like cells. Fibroblasts-like cells were observed under microscope in a condition controlled chamber for 12 h. The arrow head in (A–F) showed a representative cell with obvious movement. (GIF 27 kb)

High resolution (TIFF 2201 kb)

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Ji, S., Wang, X., Shu, J. et al. In vitro generation of myofibroblasts-like cells from liver epithelial progenitor cells of rhesus monkey (Macaca mulatta). In Vitro Cell.Dev.Biol.-Animal 47, 383–390 (2011). https://doi.org/10.1007/s11626-011-9401-z

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