Li Xiaoqin, Hayashi Ryuhei, Imaizumi Tsutomu, Harrington Jodie, Kudo Yuji, Takayanagi Hiroshi, Baba Koichi, Nishida Kohji
Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
Department of Stem Cells and Applied Medicine, Osaka University Graduate School of Medicine, Suita, Osaka, Japan.
PLoS One. 2025 Apr 21;20(4):e0321579. doi: 10.1371/journal.pone.0321579. eCollection 2025.
Corneal diseases are a leading cause of visual impairment, and their treatment remains challenging. Corneal epithelial stem cells exist in the limbus, the peripheral region of the cornea, and play an important role in corneal regeneration. Here, we evaluated the effects of extracellular vesicles from human adipose-derived mesenchymal stem cells (AdMSC-EVs) on limbal epithelial cells (LECs). Colony formation assays showed that the colony-forming efficiency of LECs significantly increased in the presence of AdMSC-EVs. We next demonstrated that AdMSC-EVs accelerated the migration of LECs in a scratch assay, whereas the proliferation of LECs was decreased by AdMSC-EVs in the cell proliferation assay. RNA sequencing analysis of LECs indicated that AdMSC-EVs maintained their stem cell properties and improved epithelial-mesenchymal transition (EMT). Furthermore, after identifying the six most abundant microRNAs (miRNAs) in AdMSC-EVs, LEC transfection with miRNA mimics indicated that miR-25, miR-191, and miR-335 were the most probable miRNA factors within AdMSC-EVs at improving colony formation ability and EMT. Taken together, our findings indicated that AdMSC-EVs enhanced the colony formation ability and EMT of LECs, and the effects of AdMSC-EVs were in-part mediated by the miRNAs within the AdMSC-EVs.
角膜疾病是视力损害的主要原因，其治疗仍然具有挑战性。角膜上皮干细胞存在于角膜边缘（角膜的周边区域），并在角膜再生中发挥重要作用。在此，我们评估了人脂肪来源间充质干细胞外泌体（AdMSC-EVs）对角膜缘上皮细胞（LECs）的影响。集落形成试验表明，在AdMSC-EVs存在的情况下，LECs的集落形成效率显著提高。接下来，我们在划痕试验中证明AdMSC-EVs加速了LECs的迁移，而在细胞增殖试验中AdMSC-EVs降低了LECs的增殖。对LECs的RNA测序分析表明，AdMSC-EVs维持了它们的干细胞特性并改善了上皮-间质转化（EMT）。此外，在鉴定出AdMSC-EVs中六种最丰富的微小RNA（miRNAs）后，用miRNA模拟物转染LECs表明，miR-25、miR-191和miR-335是AdMSC-EVs中最有可能改善集落形成能力和EMT的miRNA因子。综上所述，我们的研究结果表明，AdMSC-EVs增强了LECs的集落形成能力和EMT，且AdMSC-EVs的作用部分是由AdMSC-EVs内的miRNAs介导的。
