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脂肪来源间充质干细胞的分泌组可减弱人角膜上皮中的上皮-间质转化。

The secretome of adipose-derived mesenchymal stem cells attenuates epithelial-mesenchymal transition in human corneal epithelium.

作者信息

Shibata Shun, Hayashi Ryuhei, Okubo Toru, Kudo Yuji, Baba Koichi, Honma Yoichi, Nishida Kohji

机构信息

Department of Stem Cells and Applied Medicine, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.

Research and Development Division, ROHTO Pharmaceutical Co., Ltd., Osaka 544-8666, Japan.

出版信息

Regen Ther. 2019 Jul 2;11:114-122. doi: 10.1016/j.reth.2019.06.005. eCollection 2019 Dec.

Abstract

INTRODUCTION

Epithelial-mesenchymal transition (EMT) induces the loss of cell-cell interactions in polarized epithelial cells and converts these cells to invasive mesenchymal-like cells. It is also involved in tissue fibrosis including that occurring in some ocular surface diseases such as pterygium and in subepithelial corneal fibrosis in limbal stem cell deficiency. Here, we examined the effects of the secretome of human adipose-derived mesenchymal stem cells (AdMSCs) on EMT in human corneal epithelial cells (CECs).

METHODS

EMT was induced with transforming growth factor-β (TGF-β) in primary human CECs isolated from the human corneal limbus. The effects of the AdMSC secretome on EMT in these cells or stratified CEC sheets were analyzed by co-cultivation experiments with the addition of AdMSC conditioned-medium. The expression of EMT-related genes and proteins in CECs was analyzed. The superstructure of CECs was observed by scanning electron microscopy. Furthermore, the barrier function of CEC sheets was analyzed by measuring transepithelial electrical resistance (TER).

RESULTS

The AdMSC secretome was found to suppress EMT-related gene expression and attenuate TGF-β-induced corneal epithelial dysfunction including the dissociation of cell-cell interactions and decreases in TER in constructed CEC sheets.

CONCLUSIONS

The secretome of AdMSCs can inhibit TGF-β-induced EMT in CECs. These findings suggest that this could be a useful source for the treatment for EMT-related ocular surface diseases.

摘要

引言

上皮-间质转化(EMT)导致极化上皮细胞中细胞间相互作用丧失,并将这些细胞转化为具有侵袭性的间充质样细胞。它还参与组织纤维化,包括在某些眼表疾病(如翼状胬肉)以及角膜缘干细胞缺乏症的上皮下角膜纤维化中发生的纤维化。在此,我们研究了人脂肪来源间充质干细胞(AdMSCs)的分泌组对人角膜上皮细胞(CECs)中EMT的影响。

方法

在从人角膜缘分离的原代人CECs中用转化生长因子-β(TGF-β)诱导EMT。通过添加AdMSC条件培养基的共培养实验分析AdMSC分泌组对这些细胞或分层CEC片层中EMT的影响。分析CECs中EMT相关基因和蛋白质的表达。通过扫描电子显微镜观察CECs的超微结构。此外,通过测量跨上皮电阻(TER)分析CEC片层的屏障功能。

结果

发现AdMSC分泌组可抑制EMT相关基因表达,并减轻TGF-β诱导的角膜上皮功能障碍,包括细胞间相互作用的解离以及构建的CEC片层中TER的降低。

结论

AdMSCs的分泌组可抑制TGF-β诱导的CECs中的EMT。这些发现表明,这可能是治疗与EMT相关的眼表疾病的有用来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d1d/6609787/e4d2d93f35a7/gr1.jpg

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