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Optimization of Storage Temperature for Cultured ARPE-19 Cells

Purpose. The establishment of future retinal pigment epithelium (RPE) replacement therapy is partly dependent on the availability of tissue-engineered RPE cells, which may be enhanced by the development of suitable storage methods for RPE. This study investigates the effect of different storage temp...

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Autores principales: Pasovic, Lara, Utheim, Tor Paaske, Maria, Rima, Lyberg, Torstein, Messelt, Edward B., Aabel, Peder, Chen, Dong Feng, Chen, Xiangjun, Eidet, Jon Roger
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3819763/
https://www.ncbi.nlm.nih.gov/pubmed/24251032
http://dx.doi.org/10.1155/2013/216359
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author Pasovic, Lara
Utheim, Tor Paaske
Maria, Rima
Lyberg, Torstein
Messelt, Edward B.
Aabel, Peder
Chen, Dong Feng
Chen, Xiangjun
Eidet, Jon Roger
author_facet Pasovic, Lara
Utheim, Tor Paaske
Maria, Rima
Lyberg, Torstein
Messelt, Edward B.
Aabel, Peder
Chen, Dong Feng
Chen, Xiangjun
Eidet, Jon Roger
author_sort Pasovic, Lara
collection PubMed
description Purpose. The establishment of future retinal pigment epithelium (RPE) replacement therapy is partly dependent on the availability of tissue-engineered RPE cells, which may be enhanced by the development of suitable storage methods for RPE. This study investigates the effect of different storage temperatures on the viability, morphology, and phenotype of cultured RPE. Methods. ARPE-19 cells were cultured under standard conditions and stored in HEPES-buffered MEM at nine temperatures (4°C, 8°C, 12°C, 16°C, 20°C, 24°C, 28°C, 32°C, and 37°C) for seven days. Viability and phenotype were assessed by a microplate fluorometer and epifluorescence microscopy, while morphology was analyzed by scanning electron microscopy. Results. The percentage of viable cells preserved after storage was highest in the 16°C group (48.7% ± 9.8%; P < 0.01 compared to 4°C, 8°C, and 24°C–37°C; P < 0.05 compared to 12°C). Ultrastructure was best preserved at 12°C, 16°C, and 20°C. Expression of actin, ZO-1, PCNA, caspase-3, and RPE65 was maintained after storage at 16°C compared to control cells that were not stored. Conclusion. Out of nine temperatures tested between 4°C and 37°C, storage at 12°C, 16°C, and 20°C was optimal for maintenance of RPE cell viability, morphology, and phenotype. The preservation of RPE cells is critically dependent on storage temperature.
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spelling pubmed-38197632013-11-18 Optimization of Storage Temperature for Cultured ARPE-19 Cells Pasovic, Lara Utheim, Tor Paaske Maria, Rima Lyberg, Torstein Messelt, Edward B. Aabel, Peder Chen, Dong Feng Chen, Xiangjun Eidet, Jon Roger J Ophthalmol Research Article Purpose. The establishment of future retinal pigment epithelium (RPE) replacement therapy is partly dependent on the availability of tissue-engineered RPE cells, which may be enhanced by the development of suitable storage methods for RPE. This study investigates the effect of different storage temperatures on the viability, morphology, and phenotype of cultured RPE. Methods. ARPE-19 cells were cultured under standard conditions and stored in HEPES-buffered MEM at nine temperatures (4°C, 8°C, 12°C, 16°C, 20°C, 24°C, 28°C, 32°C, and 37°C) for seven days. Viability and phenotype were assessed by a microplate fluorometer and epifluorescence microscopy, while morphology was analyzed by scanning electron microscopy. Results. The percentage of viable cells preserved after storage was highest in the 16°C group (48.7% ± 9.8%; P < 0.01 compared to 4°C, 8°C, and 24°C–37°C; P < 0.05 compared to 12°C). Ultrastructure was best preserved at 12°C, 16°C, and 20°C. Expression of actin, ZO-1, PCNA, caspase-3, and RPE65 was maintained after storage at 16°C compared to control cells that were not stored. Conclusion. Out of nine temperatures tested between 4°C and 37°C, storage at 12°C, 16°C, and 20°C was optimal for maintenance of RPE cell viability, morphology, and phenotype. The preservation of RPE cells is critically dependent on storage temperature. Hindawi Publishing Corporation 2013 2013-10-22 /pmc/articles/PMC3819763/ /pubmed/24251032 http://dx.doi.org/10.1155/2013/216359 Text en Copyright © 2013 Lara Pasovic et al. https://creativecommons.org/licenses/by/3.0/ This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Pasovic, Lara
Utheim, Tor Paaske
Maria, Rima
Lyberg, Torstein
Messelt, Edward B.
Aabel, Peder
Chen, Dong Feng
Chen, Xiangjun
Eidet, Jon Roger
Optimization of Storage Temperature for Cultured ARPE-19 Cells
title Optimization of Storage Temperature for Cultured ARPE-19 Cells
title_full Optimization of Storage Temperature for Cultured ARPE-19 Cells
title_fullStr Optimization of Storage Temperature for Cultured ARPE-19 Cells
title_full_unstemmed Optimization of Storage Temperature for Cultured ARPE-19 Cells
title_short Optimization of Storage Temperature for Cultured ARPE-19 Cells
title_sort optimization of storage temperature for cultured arpe-19 cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3819763/
https://www.ncbi.nlm.nih.gov/pubmed/24251032
http://dx.doi.org/10.1155/2013/216359
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