Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction
Corneal endothelial cells (CECs) do not proliferate or recover after illness or injury, resulting in decreased cell density and loss of pump/barrier function. Considering the shortage of donor cornea, it is vital to establish robust methods to generate CECs from induced pluripotent stem cells (iPSCs...
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| Format: | Article |
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MDPI AG
2022-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/1/701 |
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| author | Seongjun So Yoonkyung Park Soon Suk Kang Jongsuk Han Jeong Hye Sunwoo Whanseo Lee Jin Kim Eun Ah Ye Jae Yong Kim Hungwon Tchah Eunju Kang Hun Lee |
| author_facet | Seongjun So Yoonkyung Park Soon Suk Kang Jongsuk Han Jeong Hye Sunwoo Whanseo Lee Jin Kim Eun Ah Ye Jae Yong Kim Hungwon Tchah Eunju Kang Hun Lee |
| author_sort | Seongjun So |
| collection | DOAJ |
| description | Corneal endothelial cells (CECs) do not proliferate or recover after illness or injury, resulting in decreased cell density and loss of pump/barrier function. Considering the shortage of donor cornea, it is vital to establish robust methods to generate CECs from induced pluripotent stem cells (iPSCs). We investigated the efficacy and safety of transplantation of iPSC-derived CECs into a corneal endothelial dysfunction (CED) rabbit model. iPSCs were generated from human fibroblasts. We characterized iPSCs by demonstrating the gene expression of the PSC markers <i>OCT4</i>, <i>SOX2</i>, <i>TRA-1-60</i>, and <i>NANOG</i>, teratoma formation, and differentiation into three germ layers. Differentiation of iPSCs into CECs was induced via neural crest cell (NCC) induction. CEC markers were detected using immunofluorescence and gene expression was analyzed using quantitative real-time PCR (qRT-PCR). After culturing iPSC-derived NCCs, we found the expression of zona occludens-1 (ZO-1) and Na<sup>+</sup>/K<sup>+</sup> ATPase and a hexagonal morphology. <i>ATP1A1</i>, <i>COL8A1</i>, and <i>AQP1</i> mRNA expression was higher in iPSC-derived CECs than in iPSCs and NCCs. We performed an injection of iPSC-derived CECs into the anterior chamber of a CED rabbit model and found improved levels of corneal transparency. We also found increased numbers of ZO-1- and ATP1A1-positive cells in rabbit corneas in the iPSC-derived CEC transplantation group. Usage of the coating material vitronectin (VTN) and fasudil resulted in good levels of CEC marker expression, demonstrated with Western blotting and immunocytochemistry. Combination of the VTN coating material and fasudil, instead of FNC mixture and Y27632, afforded the best results in terms of CEC differentiation’s in vitro and in vivo efficacy. Successful transplantation of CEC-like cells into a CED animal model confirms the therapeutic efficacy of these cells, demonstrated by the restoration of corneal clarity. Our results suggest that iPSC-derived CECs can be a promising cellular resource for the treatment of CED. |
| first_indexed | 2024-03-11T09:57:49Z |
| format | Article |
| id | doaj.art-5e58628ec0b248a98fcf167536e3d748 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T09:57:49Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-5e58628ec0b248a98fcf167536e3d7482023-11-16T15:37:31ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0124170110.3390/ijms24010701Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial DysfunctionSeongjun So0Yoonkyung Park1Soon Suk Kang2Jongsuk Han3Jeong Hye Sunwoo4Whanseo Lee5Jin Kim6Eun Ah Ye7Jae Yong Kim8Hungwon Tchah9Eunju Kang10Hun Lee11Department of Biomedical Science, CHA Advanced Research Institute, College of Life Science and Center for Embryo and Stem Cell Research, CHA University, Seongnam 13488, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Biomedical Science, CHA Advanced Research Institute, College of Life Science and Center for Embryo and Stem Cell Research, CHA University, Seongnam 13488, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaDepartment of Biomedical Science, CHA Advanced Research Institute, College of Life Science and Center for Embryo and Stem Cell Research, CHA University, Seongnam 13488, Republic of KoreaDepartment of Ophthalmology, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of KoreaCorneal endothelial cells (CECs) do not proliferate or recover after illness or injury, resulting in decreased cell density and loss of pump/barrier function. Considering the shortage of donor cornea, it is vital to establish robust methods to generate CECs from induced pluripotent stem cells (iPSCs). We investigated the efficacy and safety of transplantation of iPSC-derived CECs into a corneal endothelial dysfunction (CED) rabbit model. iPSCs were generated from human fibroblasts. We characterized iPSCs by demonstrating the gene expression of the PSC markers <i>OCT4</i>, <i>SOX2</i>, <i>TRA-1-60</i>, and <i>NANOG</i>, teratoma formation, and differentiation into three germ layers. Differentiation of iPSCs into CECs was induced via neural crest cell (NCC) induction. CEC markers were detected using immunofluorescence and gene expression was analyzed using quantitative real-time PCR (qRT-PCR). After culturing iPSC-derived NCCs, we found the expression of zona occludens-1 (ZO-1) and Na<sup>+</sup>/K<sup>+</sup> ATPase and a hexagonal morphology. <i>ATP1A1</i>, <i>COL8A1</i>, and <i>AQP1</i> mRNA expression was higher in iPSC-derived CECs than in iPSCs and NCCs. We performed an injection of iPSC-derived CECs into the anterior chamber of a CED rabbit model and found improved levels of corneal transparency. We also found increased numbers of ZO-1- and ATP1A1-positive cells in rabbit corneas in the iPSC-derived CEC transplantation group. Usage of the coating material vitronectin (VTN) and fasudil resulted in good levels of CEC marker expression, demonstrated with Western blotting and immunocytochemistry. Combination of the VTN coating material and fasudil, instead of FNC mixture and Y27632, afforded the best results in terms of CEC differentiation’s in vitro and in vivo efficacy. Successful transplantation of CEC-like cells into a CED animal model confirms the therapeutic efficacy of these cells, demonstrated by the restoration of corneal clarity. Our results suggest that iPSC-derived CECs can be a promising cellular resource for the treatment of CED.https://www.mdpi.com/1422-0067/24/1/701induced pluripotent stem cellcorneal endothelial cellscorneal endothelial dysfunctioncell injectioncell therapy |
| spellingShingle | Seongjun So Yoonkyung Park Soon Suk Kang Jongsuk Han Jeong Hye Sunwoo Whanseo Lee Jin Kim Eun Ah Ye Jae Yong Kim Hungwon Tchah Eunju Kang Hun Lee Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction International Journal of Molecular Sciences induced pluripotent stem cell corneal endothelial cells corneal endothelial dysfunction cell injection cell therapy |
| title | Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction |
| title_full | Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction |
| title_fullStr | Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction |
| title_full_unstemmed | Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction |
| title_short | Therapeutic Potency of Induced Pluripotent Stem-Cell-Derived Corneal Endothelial-like Cells for Corneal Endothelial Dysfunction |
| title_sort | therapeutic potency of induced pluripotent stem cell derived corneal endothelial like cells for corneal endothelial dysfunction |
| topic | induced pluripotent stem cell corneal endothelial cells corneal endothelial dysfunction cell injection cell therapy |
| url | https://www.mdpi.com/1422-0067/24/1/701 |
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