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BOYOUNG JUNG, Eun-Soon Kim, Jae Yong Kim, Hungwon Tchah, Changmo Hwang; Rabbit corneal endothelial cells expansion by Rock inhibitor and mesenchymal stem cell-derived conditioned medium. Invest. Ophthalmol. Vis. Sci. 2017;58(8):1471.
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© ARVO (1962-2015); The Authors (2016-present)
Corneal endothelial cells(CECs) have limited proliferation capacity in vivo and in vitro and therefore there exists constant demand of high proliferation of CECs in vitro for endothelial keratoplasty or cornea regeneration. The purpose of this study is to explore a potent approach for increasing passage number of CEC culture in vitro.
Our main idea was the addition of Rho-associated kinase(ROCK)-inhibitor and human mesenchymal stem cell-derived conditioned medium (MSC-CM) may influence on proliferation of CECs. CECs were isolated from the cornea of the New Zealand White Rabbit. Endothelium was peel off from cornea with descemet’s membrane and isolated using 0.25% trypsin-EDTA solution. MSC-CM was prepared from umbilical cord blood-derived MSC medium and mixed with culture medium at a ratio of 1: 1. CECs were cultured and were subcultured with the ROCK-inhibitor, Y-27632 (10µM) and MSC-CM. The ability to proliferate was confirmed through cell count and CCK-8 assay, and it was confirmed that immunofluorescent staining was performed to maintain the function of CECs.
Treatment of Y-27632 in CEC culture medium resulted in no significant effect on cell proliferation. CECs in Y-27632 treated group maintained cell morphology higher passage number and cell viability was higher during subculture than control group. MSC-CM treatment showed a 2.5-fold increase in proliferation, but when treated with Y-27632 it was rather reduced.
ROCK-inhibitor did not enhance cell proliferation but is important for maintaining function and MSC-CM enhanced cell proliferation. However, further studies are needed to find optimized in vitro culture condition for the growth of CECs.
This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.
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