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

In Vitro Cellular and Developmental Biology - Animal, ISSN: 1071-2690, Vol: 47, Issue: 5-6, Page: 383-390
2011
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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 transcriptionpolymerase 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 ?SMA, 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. © 2011 The Society for In Vitro Biology.

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