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

Document Type

Journal Article

Publication Title

Ocular Surface

Publisher

Elsevier

School

School of Medical and Health Sciences

RAS ID

34033

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

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.

DOI

10.1016/j.jtos.2019.10.009

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