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Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells

Limbal stem cell (LSC) deficiency causes progressive loss of vision but may be treated by transplant of autologous LSCs. Cryopreservation has the potential to indefinitely extend the lifespan of LSCs allowing re-transplant in case of graft failure. In this study, we aimed to identify the optimal cry...

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Autores principales: Osei-Bempong, Charles, Ghareeb, Ali E., Lako, Majlinda, Figueiredo, Francisco C., Armitage, W. John
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6167250/
https://www.ncbi.nlm.nih.gov/pubmed/30075110
http://dx.doi.org/10.1016/j.cryobiol.2018.07.008
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author Osei-Bempong, Charles
Ghareeb, Ali E.
Lako, Majlinda
Figueiredo, Francisco C.
Armitage, W. John
author_facet Osei-Bempong, Charles
Ghareeb, Ali E.
Lako, Majlinda
Figueiredo, Francisco C.
Armitage, W. John
author_sort Osei-Bempong, Charles
collection PubMed
description Limbal stem cell (LSC) deficiency causes progressive loss of vision but may be treated by transplant of autologous LSCs. Cryopreservation has the potential to indefinitely extend the lifespan of LSCs allowing re-transplant in case of graft failure. In this study, we aimed to identify the optimal cryoprotectant and cryoprotectant concentration for LSC cultures. Suspension cultures derived from cadaveric corneoscleral rims were cooled to 4 °C with Me(2)SO, propylene glycol or ethylene glycol at a concentration of 5%, 10% or 15%. Cell tolerance was measured in terms of membrane integrity, colony-forming efficiency and alamarBlue(®) reduction. Increasing cryoprotectant concentration above 5% reduced membrane integrity, metabolism and colony-forming efficiency. Cryoprotectant choice did not significantly influence these characteristics. Cells demonstrating Side Population were maintained after cryopreservation with 5% propylene glycol in vapour phase liquid nitrogen for 1 week, indicating that cryopreservation of LSCs with relatively low cryoprotectant concentration (5%) has promise in low-temperature eye banking.
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spelling pubmed-61672502018-10-03 Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells Osei-Bempong, Charles Ghareeb, Ali E. Lako, Majlinda Figueiredo, Francisco C. Armitage, W. John Cryobiology Article Limbal stem cell (LSC) deficiency causes progressive loss of vision but may be treated by transplant of autologous LSCs. Cryopreservation has the potential to indefinitely extend the lifespan of LSCs allowing re-transplant in case of graft failure. In this study, we aimed to identify the optimal cryoprotectant and cryoprotectant concentration for LSC cultures. Suspension cultures derived from cadaveric corneoscleral rims were cooled to 4 °C with Me(2)SO, propylene glycol or ethylene glycol at a concentration of 5%, 10% or 15%. Cell tolerance was measured in terms of membrane integrity, colony-forming efficiency and alamarBlue(®) reduction. Increasing cryoprotectant concentration above 5% reduced membrane integrity, metabolism and colony-forming efficiency. Cryoprotectant choice did not significantly influence these characteristics. Cells demonstrating Side Population were maintained after cryopreservation with 5% propylene glycol in vapour phase liquid nitrogen for 1 week, indicating that cryopreservation of LSCs with relatively low cryoprotectant concentration (5%) has promise in low-temperature eye banking. Elsevier 2018-10 /pmc/articles/PMC6167250/ /pubmed/30075110 http://dx.doi.org/10.1016/j.cryobiol.2018.07.008 Text en Crown Copyright © Published by Elsevier Inc. All rights reserved. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Osei-Bempong, Charles
Ghareeb, Ali E.
Lako, Majlinda
Figueiredo, Francisco C.
Armitage, W. John
Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells
title Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells
title_full Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells
title_fullStr Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells
title_full_unstemmed Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells
title_short Defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells
title_sort defining the optimal cryoprotectant and concentration for cryopreservation of limbal stem cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6167250/
https://www.ncbi.nlm.nih.gov/pubmed/30075110
http://dx.doi.org/10.1016/j.cryobiol.2018.07.008
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