Artikel
Towards experimental stem cell-based therapy for Age-related Macular Degeneration – Transplantation of patient derived-tissues in the rat subretinal space
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Autoren
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Céline Koster
- Department of Clinical Genetics, Amsterdam University Medical Centers (AUMC), University of Amsterdam (UvA), location Meibergdreef, Amsterdam/NL
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F. Barone
- Unit on Ocular Stem Cell and Translational Research, National Eye Institute (NEI), Bethesda/USA
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A. Maminishkis
- Section on Epithelial and Retinal Physiology and Disease, NEI, Bethesda/USA
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M. Jae Song
- Unit on Ocular Stem Cell and Translational Research, National Eye Institute (NEI), Bethesda/USA
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R. Quinn
- Unit on Ocular Stem Cell and Translational Research, National Eye Institute (NEI), Bethesda/USA
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C. Lewallen
- Georgia Institute of Technology, G.W. Woodruff School of Mechanical Engineering, Atlanta/USA
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J.C. van Meurs
- Rotterdam Eye Hospital, Rotterdam/NL; Department of Ophthalmology, Erasmus Medical Center, Rotterdam/NL
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F.D. Verbraak
- Department of Ophthalmology, AMC, UvA, Amsterdam/NL
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T. H. Smit
- Anatomy, Embryology and Physiology, Amsterdam Movement Sciences, AUMC, UvA, Amsterdam/NL
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J. B. ten Brink
- Department of Clinical Genetics, Amsterdam University Medical Centers (AUMC), University of Amsterdam (UvA), location Meibergdreef, Amsterdam/NL
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A.L.M.A ten Asbroek
- Department of Clinical Genetics, Amsterdam University Medical Centers (AUMC), University of Amsterdam (UvA), location Meibergdreef, Amsterdam/NL
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C.J. Boon
- Department of Ophthalmology, AMC, UvA, Amsterdam/NL; Department of Ophthalmology, Leiden University Medical Center, Leiden/NL
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K. Bharti
- Unit on Ocular Stem Cell and Translational Research, National Eye Institute (NEI), Bethesda/USA
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A.A. Bergen
- Department of Clinical Genetics, Amsterdam University Medical Centers (AUMC), University of Amsterdam (UvA), location Meibergdreef, Amsterdam/NL; Department of Ophthalmology, AMC, UvA, Amsterdam/NL; The Netherlands Institute for Neuroscience (NIN-KNAW), Amsterdam/NL
| Veröffentlicht: | 5. Februar 2020 |
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Gliederung
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Purpose: Age-related macular degeneration (AMD) is the principle cause of severe, progressive and irreversible visual impairment among the elderly. There is currently no treatment to restore vision when the function of the macula is strongly decreased. The retinal pigment epithelium (RPE) and the choroid (Ch) play major roles in the etiology of AMD. In this study we aim to transfer human induced pluripotent stem cell-derived (hiPSC) tissue into the rat subretinal space.
Methods: iPSCs were obtained from representative AMD patients and differentiated into RPE cells in vitro on an artificial Bruch’s membrane (BM; scaffolds). iPSC-RPE cells were co-cultured with hiPSC-endothelial cells, fibroblasts and pericytes to create a 3D multilayerd tissue. Tissues were implanted after in vitro maturation into the subretinal space of healthy rats and rats treated with sodium iodate. The animals were followed over time non-invasively using Electroretinography (ERG), Scanning Laser Ophthalmoscopy (SLO) and Optical Coherence Tomography (OCT) for two months. ERG responses were analyzed using their a- and b-wave amplitudes and latencies (MATLAB). At various time points, eyes were collected for histology.
Results: hiPSCs were successfully adhered and differentiated into RPE on electrospun scaffolds. These cells showed the expression of several specific RPE markers (RPE65, BEST1, MITF, MERTK, ZO-1 and others). The cells were characterized in vitro and successfully transplanted as 1 mm discs into the subretinal space of rats. SLO-OCT and Fluorescein Angiography (FA) analyses showed survival of the transplant and signs of integration. According to histology, transplants survived for at least two months in the subretinal space. ERG analyses showed reduced responses two weeks after surgery, but this was regardless of treatment.
Conclusions: We developed a pre-clinical research pipe-line, in which we are able to differentiate patient derived iPSCs to RPE and transplant these cells (with or without artificial Ch) into suitable animal models. We determined the functional and structural changes following surgery over time, non-invasively. Transplantation of (3D) RPE(-Ch) tissue into the subretinal space of an animal model opens the possibility of developing tissue therapies for retinal degenerative diseases.
