Ophthalmic Center, The Second Affiliated Hospital of Guangzhou Medical University , Guangzhou, China.
Department of Ophthalmology, The Sixth Affiliated Hospital of Guangzhou Medical University , Qingyuan, China.
Curr Eye Res. 2021 Feb;46(2):159-167. doi: 10.1080/02713683.2020.1792508. Epub 2020 Jul 13.
Human amniotic epithelial cells (HAECs) have regenerative properties and low immunogenicity, which have enabled their use without immune rejection in regenerative medicine applications, such as wound repair, corneal surgery and burn repair. The aim of this study was to explore the potential role of HAECs in the proliferation of human corneal endothelial cells (HCEnCs) and the possible mechanism of regulation.
HAECs and HCEnCs were isolated from donated tissue samples and were cultured; the collected HAEC culture medium (HAEC-Me) was added to the human corneal endothelium medium (CEM) to establish the HAEC-CM system. HCEnCs were cultured in CEM, 20%HAEC-Me, 20% HAEC-CM, 20% HAEC-CM supplemented with a GSK-3β inhibitor TWS119 or CEM supplemented with TWS119. Then, cell proliferation, apoptosis, cell cycle progression, telomerase activity, and Wnt/β-catenin pathway-related protein levels were assessed.
We found that the HCEnCs cultured in the 20% HAEC-CM had increased proliferative capacity, telomerase activity and β-catenin and Tcf4 expression levels, and they had a decrease in the rate of apoptosis and α-SMA expression when they were compared with the HCEnCs cultured in the 20% HAEC-Me. After GSK-3β was inhibited by TWS119, HCEnCs cultured in CEM or 20% HAEC-CM had an increased proliferative capacity, telomerase activity, β-catenin/Tcf4 expression and a decreased α-SMA expression, and they had a decreased apoptotic rate.
These data indicate that the human amniotic epithelial cells microenvironment can promote the proliferation of human corneal endothelial cells, which may be related to regulating telomerase activity and epithelial-to-mesenchymal transition (EMT) via the Wnt/β-catenin pathway.
人羊膜上皮细胞(HAEC)具有再生特性和低免疫原性,使其能够在再生医学应用中使用,而不会产生免疫排斥,例如伤口修复、角膜手术和烧伤修复。本研究旨在探讨 HAEC 在人角膜内皮细胞(HCEnC)增殖中的潜在作用及其可能的调节机制。
从捐赠的组织样本中分离出 HAEC 和 HCEnC 并进行培养;收集 HAEC 培养上清液(HAEC-Me)加入人角膜内皮细胞培养基(CEM)中建立 HAEC-CM 系统。HCEnC 在 CEM、20%HAEC-Me、20%HAEC-CM、20%HAEC-CM 中培养补充 GSK-3β 抑制剂 TWS119 或 CEM 中补充 TWS119。然后评估细胞增殖、凋亡、细胞周期进程、端粒酶活性和 Wnt/β-catenin 通路相关蛋白水平。
我们发现,与在 20%HAEC-Me 中培养的 HCEnC 相比,在 20%HAEC-CM 中培养的 HCEnC 增殖能力增强,端粒酶活性和β-catenin/Tcf4 表达水平升高,凋亡率降低,α-SMA 表达水平降低。在 TWS119 抑制 GSK-3β 后,在 CEM 或 20%HAEC-CM 中培养的 HCEnC 增殖能力增强,端粒酶活性、β-catenin/Tcf4 表达水平升高,α-SMA 表达水平降低,凋亡率降低。
这些数据表明,人羊膜上皮细胞微环境可以促进人角膜内皮细胞的增殖,这可能与通过 Wnt/β-catenin 通路调节端粒酶活性和上皮-间充质转化(EMT)有关。