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Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin

PURPOSE: RPE cell transplantation as a potential treatment for AMD has been extensively investigated; however, in AMD, ultrastructural damage affects both the RPE and its underlying matrix support, the Bruch's membrane (BrM). An RPE monolayer supported by a surrogate scaffold could thus provide...

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Autores principales: Galloway, Chad A., Dalvi, Sonal, Shadforth, Audra M. A., Suzuki, Shuko, Wilson, Molly, Kuai, David, Hashim, Ali, MacDonald, Leslie A., Gamm, David M., Harkin, Damien G., Singh, Ruchira
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Association for Research in Vision and Ophthalmology 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5989661/
https://www.ncbi.nlm.nih.gov/pubmed/30025113
http://dx.doi.org/10.1167/iovs.17-23157
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author Galloway, Chad A.
Dalvi, Sonal
Shadforth, Audra M. A.
Suzuki, Shuko
Wilson, Molly
Kuai, David
Hashim, Ali
MacDonald, Leslie A.
Gamm, David M.
Harkin, Damien G.
Singh, Ruchira
author_facet Galloway, Chad A.
Dalvi, Sonal
Shadforth, Audra M. A.
Suzuki, Shuko
Wilson, Molly
Kuai, David
Hashim, Ali
MacDonald, Leslie A.
Gamm, David M.
Harkin, Damien G.
Singh, Ruchira
author_sort Galloway, Chad A.
collection PubMed
description PURPOSE: RPE cell transplantation as a potential treatment for AMD has been extensively investigated; however, in AMD, ultrastructural damage affects both the RPE and its underlying matrix support, the Bruch's membrane (BrM). An RPE monolayer supported by a surrogate scaffold could thus provide a more effective approach to cell-based therapy for AMD. Toward this goal, we aimed to establish a functional human induced pluripotent stem cell–derived (hiPSC)-RPE monolayer on a Bombyx mori silk fibroin (BMSF) scaffold. METHODS: RPE differentiated from five distinct hiPSC lines were cultured on BMSF membrane coated with extracellular matrix (ECM, COL1), and either regular tissue culture plastic or Transwell coated with ECM (LAM-TCP). Morphologic, gene and protein expression, and functional characteristics of the hiPSC-RPE cultured on different membranes were compared in longitudinal experiments spanning 1 day to ≥3 months. RESULTS: The hiPSC-RPE monolayers on ECM-coated BMSF and TCP could be maintained in culture for ≥3 months and displayed RPE-characteristic morphology, pigmentation, polarity, and expression of RPE signature genes and proteins. Furthermore, hiPSC-RPE on both ECM-coated BMSF and TCP displayed robust expression and secretion of several basement membrane proteins. Importantly, hiPSC-RPE cells on COL1-BMSF and LAM-TCP showed similar efficacy in the phagocytosis and degradation of photoreceptor outer segments. CONCLUSIONS: A biomaterial scaffold manufactured from silk fibroin supports the maturation and long-term survival of a functional hiPSC-RPE monolayer. This has significant implications for both in vitro disease modeling and in vivo cell replacement therapy.
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spelling pubmed-59896612018-06-07 Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin Galloway, Chad A. Dalvi, Sonal Shadforth, Audra M. A. Suzuki, Shuko Wilson, Molly Kuai, David Hashim, Ali MacDonald, Leslie A. Gamm, David M. Harkin, Damien G. Singh, Ruchira Invest Ophthalmol Vis Sci Retinal Cell Biology PURPOSE: RPE cell transplantation as a potential treatment for AMD has been extensively investigated; however, in AMD, ultrastructural damage affects both the RPE and its underlying matrix support, the Bruch's membrane (BrM). An RPE monolayer supported by a surrogate scaffold could thus provide a more effective approach to cell-based therapy for AMD. Toward this goal, we aimed to establish a functional human induced pluripotent stem cell–derived (hiPSC)-RPE monolayer on a Bombyx mori silk fibroin (BMSF) scaffold. METHODS: RPE differentiated from five distinct hiPSC lines were cultured on BMSF membrane coated with extracellular matrix (ECM, COL1), and either regular tissue culture plastic or Transwell coated with ECM (LAM-TCP). Morphologic, gene and protein expression, and functional characteristics of the hiPSC-RPE cultured on different membranes were compared in longitudinal experiments spanning 1 day to ≥3 months. RESULTS: The hiPSC-RPE monolayers on ECM-coated BMSF and TCP could be maintained in culture for ≥3 months and displayed RPE-characteristic morphology, pigmentation, polarity, and expression of RPE signature genes and proteins. Furthermore, hiPSC-RPE on both ECM-coated BMSF and TCP displayed robust expression and secretion of several basement membrane proteins. Importantly, hiPSC-RPE cells on COL1-BMSF and LAM-TCP showed similar efficacy in the phagocytosis and degradation of photoreceptor outer segments. CONCLUSIONS: A biomaterial scaffold manufactured from silk fibroin supports the maturation and long-term survival of a functional hiPSC-RPE monolayer. This has significant implications for both in vitro disease modeling and in vivo cell replacement therapy. The Association for Research in Vision and Ophthalmology 2018-06 /pmc/articles/PMC5989661/ /pubmed/30025113 http://dx.doi.org/10.1167/iovs.17-23157 Text en Copyright 2018 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
spellingShingle Retinal Cell Biology
Galloway, Chad A.
Dalvi, Sonal
Shadforth, Audra M. A.
Suzuki, Shuko
Wilson, Molly
Kuai, David
Hashim, Ali
MacDonald, Leslie A.
Gamm, David M.
Harkin, Damien G.
Singh, Ruchira
Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin
title Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin
title_full Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin
title_fullStr Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin
title_full_unstemmed Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin
title_short Characterization of Human iPSC-RPE on a Prosthetic Bruch's Membrane Manufactured From Silk Fibroin
title_sort characterization of human ipsc-rpe on a prosthetic bruch's membrane manufactured from silk fibroin
topic Retinal Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5989661/
https://www.ncbi.nlm.nih.gov/pubmed/30025113
http://dx.doi.org/10.1167/iovs.17-23157
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