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

Lookup NU author(s): Dr Charlie Osei-Bempong, Dr Ali Ghareeb, Professor Majlinda LakoORCiD, Professor Francisco FigueiredoORCiD



This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract 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 Me2SO, 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.

Publication metadata

Author(s): Osei-Bempong C, Ghareeb AE, Lako M, Figueiredo FC, Armitage WJ

Publication type: Article

Publication status: Published

Journal: Cryobiology

Year: 2018

Volume: 84

Pages: 98-102

Print publication date: 01/10/2018

Online publication date: 31/07/2018

Acceptance date: 30/07/2018

Date deposited: 20/08/2018

ISSN (print): 0011-2240

ISSN (electronic): 1090-2392

Publisher: Academic Press


DOI: 10.1016/j.cryobiol.2018.07.008


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Funder referenceFunder name
614620European Research Council (ERC)
G0900879Medical Research Council (MRC)
MC_PC_15030Medical Research Council (MRC)