Amniotic Membrane Enhances the Characteristics and Function of Stem Cell-Derived Retinal Pigment Epithelium Sheets by Inhibiting the Epithelial-Mesenchymal Transition.
Amniotic Membrane Enhances the Characteristics and Function of Stem Cell-Derived Retinal Pigment Epithelium Sheets by Inhibiting the Epithelial-Mesenchymal Transition.
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DOI：
10.1016/j.actbio.2022.07.064
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发表时间：
2022-08
影响因子：
9.7
通讯作者：
Suai Zhang;Ke Ye;Guanjie Gao;Xiaojing Song;P. Xu;Jingrong Zeng;Bingbing Xie;Dandan Zheng;
中科院分区：
文献类型：
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作者：
Suai Zhang;Ke Ye;Guanjie Gao;Xiaojing Song;P. Xu;Jingrong Zeng;Bingbing Xie;Dandan Zheng;
人多能干细胞衍生的视网膜色素上皮（iRPE）是用于具有RPE缺陷的视网膜病症的疾病建模和细胞替代疗法的有吸引力的细胞来源。然而，仍然存在挑战，以开发接近体内微环境的适当培养条件来产生iRPE片，其更忠实地模拟人RPE细胞的特征和功能。在这里，我们开发了一个简单的，新颖的平台，使用人羊膜（hAM）作为天然支架构建真实的iRPE片。脱细胞hAM（dAM）提供了Bruch膜（BM）样生物支架，支持iRPE生长并增强了iRPE细胞的上皮特征、极性分布和功能特征。重要的是，进行RNA-seq分析以比较在不同基质上培养的iRPE细胞的转录组，这揭示了dAM支持和促进iRPE生长的潜在机制是抑制上皮间质转化（EMT）。组织工程化iRPE片移植到兔视网膜下腔后，细胞存活并保持单层。总之，我们的结果表明，模仿天然BM的dAM允许工程化真实的人类RPE片，这将为视网膜退行性疾病的疾病建模、药物筛选和细胞替代治疗提供有价值的生物材料。工程化的RPE膜比用于移植的RPE细胞悬液具有更大的优势，因为它们支持RPE在完整的单层中生长，而RPE功能依赖于RPE细胞的单层生长。RPE培养的基质对于维持干细胞中RPE的生理功能起着关键作用视网膜变性患者的细胞疗法。在这项研究中，我们构建了工程iRPE片脱细胞的人羊膜（dAM）的支架，这有助于提高上皮细胞的特性，极性分布和功能特性的iRPE。dAM表现出抗上皮间质转化（EMT）支持iRPE生长的能力。体内移植的结果进一步证实了iRPE膜在视网膜再生治疗中的可行性。在dAM上工程化RPE片材是促进iRPE替代疗法和视网膜疾病建模的发展的有前景的策略。
Human pluripotent stem cell-derived retinal pigment epithelium (iRPE) is an attractive cell source for disease modeling and cell replacement therapy of retinal disorders with RPE defects. However, there are still challenges to develop appropriate culture conditions close toin vivomicroenvironment to generate iRPE sheets, which mimic more faithfully the characteristics and functions of the human RPE cells. Here, we developed a simple, novel platform to construct authentic iRPE sheets using human amniotic membrane (hAM) as a natural scaffold. The decellularized hAM (dAM) provided a Bruch's membrane (BM)-like bioscaffold, supported the iRPE growth and enhanced the epithelial features, polarity distribution and functional features of iRPE cells. Importantly, RNA-seq analysis was performed to compare the transcriptomes of iRPE cells cultured on different substrates, which revealed the potential mechanism that dAM supported and promoted iRPE growth was the inhibition of epithelial mesenchymal transition (EMT). The tissue-engineered iRPE sheets survived and kept monolayer when transplanted into the subretinal space of rabbits. All together, our results indicate that the dAM imitating the natural BM allows for engineering authentic human RPE sheets, which will provide valuable biomaterials for disease modeling, drug screening and cell replacement therapy of retinal degenerative diseases.Statement of significance:Engineered RPE sheets have a great advantage over RPE cell suspension for transplantation as they support RPE growth in an intact monolayer which RPE functions are dependent on. The substrates for RPE culture play a critical role to maintain the physiological functions of the RPE in stem cell therapies for patients with retinal degeneration. In this study, we constructed engineered iRPE sheets on the decellularized human amniotic membrane (dAM) scaffolds, which contributed to enhancing epithelial features, polarity distribution and functional features of iRPE. dAM exhibited the ability of anti-epithelial mesenchymal transition (EMT) to support iRPE growth. Furtherly, the results of transplantedin vivodemonstrated the feasibility of iRPE sheets in retina regenerative therapy. Engineering RPE sheets on dAM is a promising strategy to facilitate the development of iRPE replacement therapy and retinal disease modeling.