Investigation of factors associated with ABCB5-positive limbal stem cell isolation yields from human donors

Abstract

Purpose: To identify factors associated with isolation yields of ATP-binding cassette (ABC) superfamily member B5 (ABCB5)-positive limbal stem cells (LSCs) from human cadaveric donor eyes. Methods: Whole eye globes were obtained from the Saving Sight eye bank, Kansas City, MO and the CorneaGen eye bank, Seattle, WA. ABCB5-positive LSCs were sorted by flow cytometry upon anti-ABCB5 monoclonal antibody staining within one week after donor death. The yields of live limbal epithelial cells in their entirety and of isolated pure ABCB5-positive LSC subsets were correlated with variables contained in the eye donors’ medical information. Results: The mean isolation yield of live limbal epithelial cells and ABCB5-positive LSCs per donor eye was (340,000 ± 160,000 and 2,608 ± 1,842 respectively, mean ± SD). Stepwise regression analysis showed that cardiac disease-related death was the strongest negative predictor of the ABCB5-positive LSC isolation yield (p = 0.01). While we observed a trend for an age-related decline in the yield of ABCB5-positive LSCs, a statistically significant association could not be established (2% decrease/year, p = 0.11). Additionally, despite a trend for decreased isolation yields of total live limbal epithelial cells isolated from single donors with a longer time between death and tissue processing (p = 0.04), this did not affect the yields of purified ABCB5-positive LSC, which was independent of increasing time between death and tissue processing (p = 0.50). Conclusions: Our study identifies cardiac disease-related death as a donor variable significantly associated with lower ABCB5-positive LSC isolation yields.

RAS ID

34033

Document Type

Journal Article

Date of Publication

1-1-2020

School

School of Medical and Health Sciences

Copyright

subscription content

Publisher

Elsevier

Comments

Sasamoto, Y., Sasamoto, N., Tran, J., Mishra, A., Ksander, B. R., Frank, M. H., & Frank, N. Y. (2020). Investigation of factors associated with ABCB5-positive limbal stem cell isolation yields from human donors. The Ocular Surface, 18, 114-120. https://doi.org/10.1016/j.jtos.2019.10.009

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Link to publisher version (DOI)

10.1016/j.jtos.2019.10.009