Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse Retina
Cell replacement therapies may be key in achieving functional recovery in neurodegenerative optic neuropathies diseases such as glaucoma. One strategy that holds promise in this regard is the use of human embryonic stem cell and induced pluripotent stem-derived retinal ganglion cells (hRGCs). Previo...
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MDPI AG
2022-10-01
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Online Access: | https://www.mdpi.com/2073-4409/11/20/3241 |
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author | Louis-Philippe Croteau Michael L. Risner Lauren K. Wareham Nolan R. McGrady Xitiz Chamling Donald J. Zack David J. Calkins |
author_facet | Louis-Philippe Croteau Michael L. Risner Lauren K. Wareham Nolan R. McGrady Xitiz Chamling Donald J. Zack David J. Calkins |
author_sort | Louis-Philippe Croteau |
collection | DOAJ |
description | Cell replacement therapies may be key in achieving functional recovery in neurodegenerative optic neuropathies diseases such as glaucoma. One strategy that holds promise in this regard is the use of human embryonic stem cell and induced pluripotent stem-derived retinal ganglion cells (hRGCs). Previous hRGC transplantation studies have shown modest success. This is in part due to the low survival and integration of the transplanted cells in the host retina. The field is further challenged by mixed assays and outcome measurements that probe and determine transplantation success. Thefore, we have devised a transplantation assay involving hRGCs and mouse retina explants that bypasses physical barriers imposed by retinal membranes. We show that hRGC neurites and somas are capable of invading mouse explants with a subset of hRGC neurites being guided by mouse RGC axons. Neonatal mouse retina explants, and to a lesser extent, adult explants, promote hRGC integrity and neurite outgrowth. Using this assay, we tested whether suppmenting cultures with brain derived neurotrophic factor (BDNF) and the adenylate cyclase activator, forskolin, enhances hRGC neurite integration, neurite outgrowth, and integrity. We show that supplementing cultures with a combination BDNF and forskolin strongly favors hRGC integrity, increasing neurite outgrowth and complexity as well as the invasion of mouse explants. The transplantation assay presented here is a practical tool for investigating strategies for testing and optimizing the integration of donor cells into host tissues. |
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issn | 2073-4409 |
language | English |
last_indexed | 2024-03-09T20:29:10Z |
publishDate | 2022-10-01 |
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series | Cells |
spelling | doaj.art-7050f2aefc1048caa5c35aa11ef8465a2023-11-23T23:27:53ZengMDPI AGCells2073-44092022-10-011120324110.3390/cells11203241Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse RetinaLouis-Philippe Croteau0Michael L. Risner1Lauren K. Wareham2Nolan R. McGrady3Xitiz Chamling4Donald J. Zack5David J. Calkins6Department of Ophthalmology and Visual Sciences, Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN 37232, USADepartment of Ophthalmology and Visual Sciences, Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN 37232, USADepartment of Ophthalmology and Visual Sciences, Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN 37232, USADepartment of Ophthalmology and Visual Sciences, Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN 37232, USADepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USADepartment of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USADepartment of Ophthalmology and Visual Sciences, Vanderbilt Eye Institute, Vanderbilt University Medical Center, Nashville, TN 37232, USACell replacement therapies may be key in achieving functional recovery in neurodegenerative optic neuropathies diseases such as glaucoma. One strategy that holds promise in this regard is the use of human embryonic stem cell and induced pluripotent stem-derived retinal ganglion cells (hRGCs). Previous hRGC transplantation studies have shown modest success. This is in part due to the low survival and integration of the transplanted cells in the host retina. The field is further challenged by mixed assays and outcome measurements that probe and determine transplantation success. Thefore, we have devised a transplantation assay involving hRGCs and mouse retina explants that bypasses physical barriers imposed by retinal membranes. We show that hRGC neurites and somas are capable of invading mouse explants with a subset of hRGC neurites being guided by mouse RGC axons. Neonatal mouse retina explants, and to a lesser extent, adult explants, promote hRGC integrity and neurite outgrowth. Using this assay, we tested whether suppmenting cultures with brain derived neurotrophic factor (BDNF) and the adenylate cyclase activator, forskolin, enhances hRGC neurite integration, neurite outgrowth, and integrity. We show that supplementing cultures with a combination BDNF and forskolin strongly favors hRGC integrity, increasing neurite outgrowth and complexity as well as the invasion of mouse explants. The transplantation assay presented here is a practical tool for investigating strategies for testing and optimizing the integration of donor cells into host tissues.https://www.mdpi.com/2073-4409/11/20/3241human stem cellsretinal ganglion cellmouse retina explantglaucomacell replacement therapy |
spellingShingle | Louis-Philippe Croteau Michael L. Risner Lauren K. Wareham Nolan R. McGrady Xitiz Chamling Donald J. Zack David J. Calkins Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse Retina Cells human stem cells retinal ganglion cell mouse retina explant glaucoma cell replacement therapy |
title | Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse Retina |
title_full | Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse Retina |
title_fullStr | Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse Retina |
title_full_unstemmed | Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse Retina |
title_short | Ex Vivo Integration of Human Stem Retinal Ganglion Cells into the Mouse Retina |
title_sort | ex vivo integration of human stem retinal ganglion cells into the mouse retina |
topic | human stem cells retinal ganglion cell mouse retina explant glaucoma cell replacement therapy |
url | https://www.mdpi.com/2073-4409/11/20/3241 |
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