Pluripotent Stem Cell-Derived Corneal Endothelial Cells Form a Functional Corneal Endothelium in Canines.

IF 2.6 3区医学 Q2 OPHTHALMOLOGY

Translational Vision Science & Technology Pub Date : 2025-10-01 DOI:10.1167/tvst.14.10.5

Muhammad Ali, Shahid Y Khan, Hira Butt, Ryan C Navarro, Jessica M Izzi, S Amer Riazuddin

{"title":"Pluripotent Stem Cell-Derived Corneal Endothelial Cells Form a Functional Corneal Endothelium in Canines.","authors":"Muhammad Ali, Shahid Y Khan, Hira Butt, Ryan C Navarro, Jessica M Izzi, S Amer Riazuddin","doi":"10.1167/tvst.14.10.5","DOIUrl":null,"url":null,"abstract":"Purpose: We previously reported that cryopreserved human embryonic stem cell (hESC)-derived corneal endothelial cells (CECs) can form a functional corneal endothelium (CE) on denuded Descemet's membrane (DM) in rabbits and monkeys. Here, we extend the utility of cryopreserved CECs to develop a possible treatment for canines with corneal endothelial dysfunction.Methods: We developed a corneal endothelial dysfunction canine model (n = 2) by mechanical scraping of central (8 mm diameter) CE. To regenerate the CE, the canines received an injection of cryopreserved hESC-derived CECs in the anterior chamber of the eye, followed by being placed in a cornea-down position to allow the injected cells to settle on denuded DM. The structural integrity, pachymetry, CE cell density and morphology of the regenerated and the CE of the untreated eyes were examined by optical coherence tomography (OCT), and a specular microscope. Immunohistochemical (IHC) analysis of the regenerated and resident CE was completed with a human-specific nucleoli antibody and other antibodies.Results: The corneas of the cryopreserved hESC-derived CEC-injected eyes developed corneal edema within 24 hours after scraping of the CE and cell injection but regained transparency in one to two weeks after the CEC injection. OCT revealed anatomically normal corneas with pachymetry of the CEC-injected eyes similar to the untreated left eyes, and specular microscopy examination illustrated regenerated CE exhibiting hexagonal/polygonal cells with CEC density of the regenerated CE comparable to the CE of the untreated eye. IHC analysis confirmed the regeneration of a functional CE resulting from the settlement of injected cryopreserved hESC-derived CECs on the denuded DM.Conclusions: The results confirm the regeneration of a functional CE in canines resulting from the settlement of cryopreserved hESC-derived CECs on the denuded DM.Translational relevance: Our data confirm the efficacy of cryopreserved pluripotent stem cell-derived CECs as a therapeutic option to regenerate a functional CE in canines with corneal endothelial dysfunction.","PeriodicalId":23322,"journal":{"name":"Translational Vision Science & Technology","volume":"14 10","pages":"5"},"PeriodicalIF":2.6000,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12510385/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Translational Vision Science & Technology","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1167/tvst.14.10.5","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"OPHTHALMOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

Purpose: We previously reported that cryopreserved human embryonic stem cell (hESC)-derived corneal endothelial cells (CECs) can form a functional corneal endothelium (CE) on denuded Descemet's membrane (DM) in rabbits and monkeys. Here, we extend the utility of cryopreserved CECs to develop a possible treatment for canines with corneal endothelial dysfunction.

Methods: We developed a corneal endothelial dysfunction canine model (n = 2) by mechanical scraping of central (8 mm diameter) CE. To regenerate the CE, the canines received an injection of cryopreserved hESC-derived CECs in the anterior chamber of the eye, followed by being placed in a cornea-down position to allow the injected cells to settle on denuded DM. The structural integrity, pachymetry, CE cell density and morphology of the regenerated and the CE of the untreated eyes were examined by optical coherence tomography (OCT), and a specular microscope. Immunohistochemical (IHC) analysis of the regenerated and resident CE was completed with a human-specific nucleoli antibody and other antibodies.

Results: The corneas of the cryopreserved hESC-derived CEC-injected eyes developed corneal edema within 24 hours after scraping of the CE and cell injection but regained transparency in one to two weeks after the CEC injection. OCT revealed anatomically normal corneas with pachymetry of the CEC-injected eyes similar to the untreated left eyes, and specular microscopy examination illustrated regenerated CE exhibiting hexagonal/polygonal cells with CEC density of the regenerated CE comparable to the CE of the untreated eye. IHC analysis confirmed the regeneration of a functional CE resulting from the settlement of injected cryopreserved hESC-derived CECs on the denuded DM.

Conclusions: The results confirm the regeneration of a functional CE in canines resulting from the settlement of cryopreserved hESC-derived CECs on the denuded DM.

Translational relevance: Our data confirm the efficacy of cryopreserved pluripotent stem cell-derived CECs as a therapeutic option to regenerate a functional CE in canines with corneal endothelial dysfunction.

查看原文本刊更多论文

多能干细胞衍生的角膜内皮细胞在犬中形成功能性角膜内皮。

目的：我们以前报道过冷冻保存的人胚胎干细胞（hESC）来源的角膜内皮细胞（CECs）可以在兔和猴的剥去的Descemet膜（DM）上形成功能性角膜内皮（CE）。在这里，我们扩展了冷冻保存cec的效用，以开发一种治疗角膜内皮功能障碍的可能方法。方法：采用角膜中央（直径8 mm） CE机械刮拭法建立角膜内皮功能障碍犬模型（n = 2）。为了再生CE，犬在眼睛前房注射冷冻保存的hesc来源的cec，然后将注射的细胞放置在角膜下的位置，使注射的细胞能够在剥去的DM上沉淀。通过光学相干断层扫描（OCT）和反射显微镜检查再生眼和未治疗眼的CE的结构完整性、厚度、CE细胞密度和形态学。用人类特异性核仁抗体和其他抗体完成再生和固定CE的免疫组织化学（IHC）分析。结果：低温保存的hesc源性CEC注射眼角膜在刮除CE和细胞注射后24小时内出现角膜水肿，但在CEC注射后1 ~ 2周恢复透明。OCT显示解剖上正常的角膜，注射了CEC的眼睛与未治疗的左眼相似，镜面显微镜检查显示再生CE显示六边形/多边形细胞，再生CE的CEC密度与未治疗的眼睛相当。IHC分析证实，注射冷冻保存的hesc来源的CECs沉积在去角质的dm上，可以再生功能性CE。结论：结果证实，冷冻保存的hesc来源的CECs沉积在去角质的dm上，可以再生功能性CE。我们的数据证实了冷冻保存的多能干细胞来源的cec作为再生角膜内皮功能障碍犬功能CE的治疗选择的有效性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Translational Vision Science & Technology Engineering-Biomedical Engineering

CiteScore

5.70

自引率

3.30%

发文量

346

审稿时长

25 weeks

期刊介绍： Translational Vision Science & Technology (TVST), an official journal of the Association for Research in Vision and Ophthalmology (ARVO), an international organization whose purpose is to advance research worldwide into understanding the visual system and preventing, treating and curing its disorders, is an online, open access, peer-reviewed journal emphasizing multidisciplinary research that bridges the gap between basic research and clinical care. A highly qualified and diverse group of Associate Editors and Editorial Board Members is led by Editor-in-Chief Marco Zarbin, MD, PhD, FARVO. The journal covers a broad spectrum of work, including but not limited to: Applications of stem cell technology for regenerative medicine, Development of new animal models of human diseases, Tissue bioengineering, Chemical engineering to improve virus-based gene delivery, Nanotechnology for drug delivery, Design and synthesis of artificial extracellular matrices, Development of a true microsurgical operating environment, Refining data analysis algorithms to improve in vivo imaging technology, Results of Phase 1 clinical trials, Reverse translational ("bedside to bench") research. TVST seeks manuscripts from scientists and clinicians with diverse backgrounds ranging from basic chemistry to ophthalmic surgery that will advance or change the way we understand and/or treat vision-threatening diseases. TVST encourages the use of color, multimedia, hyperlinks, program code and other digital enhancements.