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Cited by 300

The DOI matches its Crossref record; 45 of the 45 references it cites were found in a registry or an index.

Enhancement on Primate Corneal Endothelial Cell Survival In Vitro by a ROCK Inhibitor

Okumura, Naoki, Ueno, Morio, Koizumi, Noriko, Sakamoto, Yuji, Hirata, Kana, Hamuro, Junji, Kinoshita, Shigeru·2009·Investigative Opthalmology & Visual Science
DOI 10.1167/iovs.08-2634
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Abstract

PURPOSE: The transplantation of cultivated corneal endothelial cells (CECs) has gained attention recently for the treatment of patients with corneal endothelial dysfunction. However, an efficient culturing technique for human (H)CECs has yet to be properly established. The present study was conducted to investigate the applicability of the Rho kinase (ROCK) inhibitor Y-27632 in promoting cultivation of cynomolgus monkey (M)CECs. METHODS: MCECs of cynomolgus monkeys were cultured in a medium containing 10 microM Y-27632. The number of viable cells adherent to culture plates were monitored by a luminescent cell-viability assay and colony growth was detected by toluidine blue staining. Proliferating cells were detected by Ki67 expression using flow cytometry and a BrdU-labeling assay for immunocytochemistry. Annexin V-positive apoptotic cells were analyzed by flow cytometry. RESULTS: The number of viable cultivated MCECs was enhanced by Y-27632 addition after 24 hours in culture. The colony area of the culture in the presence of Y-27632 was higher than in the absence of Y-27632 on day 10. In Y-27632-treated cultures, the number of Ki67-positive cells was significantly increased at 24 and 48 hours, and the number of proliferating BrdU-positive cells was increased at 48 hours. The number of Annexin V-positive apoptotic cells was decreased at 24 hours. CONCLUSIONS: The inhibition of Rho/ROCK signaling by specific ROCK inhibitor Y-27632 promoted the adhesion of MCECs, inhibited apoptosis, and increased the number of proliferating cells. These results suggest that the ROCK inhibitor may serve as a new tool for cultivating HCECs for transplantation.

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Cited by

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Metadata

Authors
Okumura, N., Ueno, M., Koizumi, N., Sakamoto, Y., Hirata, K., Hamuro, J., & Kinoshita, S.
Venue
Investigative Opthalmology & Visual Science
Publisher
Association for Research in Vision and Ophthalmology (ARVO)
Published
2009-08-01
Volume
50
Issue
8
Pages
3680
ISSN
  • 0146-0404 (ISSN-L)
  • 1552-5783
DOI
10.1167/iovs.08-2634
OpenAlex
W2080010894
Cited by
300
References
45
FWCI
5.40
Type
journal article
Language
en
Open access
closed

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