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

Charles Osei-Bempong, Ali E. Ghareeb, Majlinda Lako, Francisco C. Figueiredo, W. John Armitage

Research output: Contribution to journalArticle (Academic Journal)peer-review

5 Citations (Scopus)
215 Downloads (Pure)


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.
Original languageEnglish
Pages (from-to)98-102
Number of pages5
Early online date31 Jul 2018
Publication statusPublished - 1 Oct 2018


  • Cell tolerance
  • Cornea
  • Corneal transplant
  • Cryopreservation
  • Dimethyl sulphoxide
  • Ethylene glycol
  • Eye bank
  • Limbal stem cell deficiency
  • Limbal stem cells
  • Propylene glycol


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