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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Main Authors: Louis-Philippe Croteau, Michael L. Risner, Lauren K. Wareham, Nolan R. McGrady, Xitiz Chamling, Donald J. Zack, David J. Calkins
Format: Article
Language:English
Published: MDPI AG 2022-10-01
Series:Cells
Subjects:
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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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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