Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency
Ocular surface reconstruction is essential for treating corneal epithelial defects and vision recovery. Stem cell-based therapy demonstrates promising results but requires further research to elucidate stem cell survival, growth, and differentiation after transplantation in vivo. This study examined...
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MDPI AG
2023-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/6/5431 |
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| author | Julia I. Khorolskaya Daria A. Perepletchikova Kirill E. Zhurenkov Daniel V. Kachkin Aleksandr A. Rubel Miralda I. Blinova Natalia A. Mikhailova |
| author_facet | Julia I. Khorolskaya Daria A. Perepletchikova Kirill E. Zhurenkov Daniel V. Kachkin Aleksandr A. Rubel Miralda I. Blinova Natalia A. Mikhailova |
| author_sort | Julia I. Khorolskaya |
| collection | DOAJ |
| description | Ocular surface reconstruction is essential for treating corneal epithelial defects and vision recovery. Stem cell-based therapy demonstrates promising results but requires further research to elucidate stem cell survival, growth, and differentiation after transplantation in vivo. This study examined the corneal reconstruction promoted by EGFP-labeled limbal mesenchymal stem cells (L-MSCs-EGFP) and their fate after transplantation. EGFP labeling allowed us to evaluate the migration and survival rates of the transferred cells. L-MSCs-EGFP seeded onto decellularized human amniotic membrane (dHAM) were transplanted into rabbits with a modeled limbal stem cell deficiency. The localization and viability of the transplanted cells in animal tissue were analyzed using histology, immunohistochemistry, and confocal microscopy up to 3 months after transplantation. EGFP-labeled cells remained viable for the first 14 days after transplantation. By the 90th day, epithelialization of the rabbit corneas reached 90%, but the presence of viable labeled cells was not observed within the newly formed epithelium. Although labeled cells demonstrated low survivability in host tissue, the squamous corneal-like epithelium was partially restored by the 30th day after transplantation of the tissue-engineered graft. Overall, this study paves the way for further optimization of transplantation conditions and studying the mechanisms of corneal tissue restoration. |
| first_indexed | 2024-03-11T06:26:09Z |
| format | Article |
| id | doaj.art-79d2493d5c3e4613924f95150986505c |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-11T06:26:09Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-79d2493d5c3e4613924f95150986505c2023-11-17T11:33:50ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-03-01246543110.3390/ijms24065431Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell DeficiencyJulia I. Khorolskaya0Daria A. Perepletchikova1Kirill E. Zhurenkov2Daniel V. Kachkin3Aleksandr A. Rubel4Miralda I. Blinova5Natalia A. Mikhailova6Institute of Cytology Russian Academy of Science, 194064 St. Petersburg, RussiaInstitute of Cytology Russian Academy of Science, 194064 St. Petersburg, RussiaInstitute of Cytology Russian Academy of Science, 194064 St. Petersburg, RussiaLaboratory of Amyloid Biology, St. Petersburg State University, 199034 St. Petersburg, RussiaLaboratory of Amyloid Biology, St. Petersburg State University, 199034 St. Petersburg, RussiaInstitute of Cytology Russian Academy of Science, 194064 St. Petersburg, RussiaInstitute of Cytology Russian Academy of Science, 194064 St. Petersburg, RussiaOcular surface reconstruction is essential for treating corneal epithelial defects and vision recovery. Stem cell-based therapy demonstrates promising results but requires further research to elucidate stem cell survival, growth, and differentiation after transplantation in vivo. This study examined the corneal reconstruction promoted by EGFP-labeled limbal mesenchymal stem cells (L-MSCs-EGFP) and their fate after transplantation. EGFP labeling allowed us to evaluate the migration and survival rates of the transferred cells. L-MSCs-EGFP seeded onto decellularized human amniotic membrane (dHAM) were transplanted into rabbits with a modeled limbal stem cell deficiency. The localization and viability of the transplanted cells in animal tissue were analyzed using histology, immunohistochemistry, and confocal microscopy up to 3 months after transplantation. EGFP-labeled cells remained viable for the first 14 days after transplantation. By the 90th day, epithelialization of the rabbit corneas reached 90%, but the presence of viable labeled cells was not observed within the newly formed epithelium. Although labeled cells demonstrated low survivability in host tissue, the squamous corneal-like epithelium was partially restored by the 30th day after transplantation of the tissue-engineered graft. Overall, this study paves the way for further optimization of transplantation conditions and studying the mechanisms of corneal tissue restoration.https://www.mdpi.com/1422-0067/24/6/5431GFP-labeled cellsocular cell therapycornea regenerationregenerative ophthalmologylimbal mesenchymal stem cells |
| spellingShingle | Julia I. Khorolskaya Daria A. Perepletchikova Kirill E. Zhurenkov Daniel V. Kachkin Aleksandr A. Rubel Miralda I. Blinova Natalia A. Mikhailova Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency International Journal of Molecular Sciences GFP-labeled cells ocular cell therapy cornea regeneration regenerative ophthalmology limbal mesenchymal stem cells |
| title | Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency |
| title_full | Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency |
| title_fullStr | Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency |
| title_full_unstemmed | Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency |
| title_short | Corneal Reconstruction with EGFP-Labelled Limbal Mesenchymal Stem Cells in a Rabbit Model of Limbal Stem Cell Deficiency |
| title_sort | corneal reconstruction with egfp labelled limbal mesenchymal stem cells in a rabbit model of limbal stem cell deficiency |
| topic | GFP-labeled cells ocular cell therapy cornea regeneration regenerative ophthalmology limbal mesenchymal stem cells |
| url | https://www.mdpi.com/1422-0067/24/6/5431 |
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