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Sci Rep. 2015 Mar 16;5:9167. doi: 10.1038/srep09167.

The effects of Rho-associated kinase inhibitor Y-27632 on primary human corneal endothelial cells propagated using a dual media approach.

Author information

1
1] Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore [2] Duke-NUS Graduate Medical School, Singapore.
2
Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore.
3
1] Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore [2] Singapore National Eye Centre, Singapore [3] Yong Loo Lin School of Medicine, National University of Singapore, Singapore.
4
1] Tissue Engineering and Stem Cell Group, Singapore Eye Research Institute, Singapore [2] Duke-NUS Graduate Medical School, Singapore [3] Singapore National Eye Centre, Singapore [4] School of Material Science and Engineering, Nanyang Technological University, Singapore.

Abstract

The global shortage of donor corneas has garnered extensive interest in the development of graft alternatives suitable for endothelial keratoplasty using cultivated primary human corneal endothelial cells (CECs). We have recently described a dual media approach for the propagation of human CECs. In this work, we characterize the effects of a Rho-kinase inhibitor Y-27632 on the cultivation of CECs propagated using the dual media culture system. Seventy donor corneas deemed unsuitable for transplantation were procured for this study. We assessed the use of Y-27632 for its effect at each stage of the cell culture process, specifically for cell attachment, cell proliferation, and during both regular passaging and cryopreservation. Lastly, comparison of donor-matched CEC-cultures expanded with or without Y-27632 was also performed. Our results showed that Y-27632 significantly improved the attachment and proliferation of primary CECs. A non-significant pro-survival effect was detected during regular cellular passage when CECs were pre-treated with Y-27632, an effect that became more evident during cryopreservation. Our study showed that the inclusion of Y-27632 was beneficial for the propagation of primary CECs expanded via the dual media approach, and was able to increase overall cell yield by between 1.96 to 3.36 fold.

PMID:
25823914
PMCID:
PMC4387913
DOI:
10.1038/srep09167
[Indexed for MEDLINE]
Free PMC Article

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