J Korean Ophthalmol Soc > Volume 59(6); 2018 > Article
Journal of the Korean Ophthalmological Society 2018;59(6):497-504.
DOI: https://doi.org/10.3341/jkos.2018.59.6.497    Published online June 15, 2018.
Xenotransplantation of Cultured Human Corneal Endothelial Cell Sheets.
Yoon Pyo Lee, Tae Young Chung, Joon Young Hyon, Young Joo Shin
1Department of Ophthalmology, Kangnam Sacred Heart Hospital, Hallym University College of Medicine, Seoul, Korea. schinn@daum.net
2Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.
3Department of Ophthalmology, Seoul National University Bundang Hospital, Seongnam, Korea.
배양사람각막내피세포판의 이종이식
이윤표1⋅정태영2⋅현준영3⋅신영주1
한림대학교 의과대학 강남성심병원 안과학교실1, 성균관대학교 의과대학 삼성서울병원 안과학교실2, 분당서울대학교병원 안과3
Correspondence:  Young Joo Shin,
Email: schinn@daum.net
Received: 22 February 2018   • Revised: 16 April 2018   • Accepted: 4 June 2018
Abstract
PURPOSE
To investigate the possibility of transplantation into rabbits of collagenase-induced cultured human corneal endothelial cell plate-rabbit corneal stromal complexes. METHODS: Human corneal endothelial cells were isolated from residual corneal limbus samples and treated with collagenase to create corneal endothelial cell sheets. Cultured sheets were transplanted into the rabbit stroma after the rabbit corneal endothelial cells and Descemet's membrane were removed. Hematoxylin-and-eosin staining and immunofluorescence staining for collagen VIIIa2 (COL8A2) and zonula occludens-1 (ZO-1) were performed. The cultured human corneal endothelial cell sheet-rabbit corneal stromal complex was transplanted into rabbits. On days 3, 5, and 7, the transplanted corneas were photographed and corneal opacity was measured. One week later, the rabbits were sacrificed. Hematoxylin-and-eosin staining and immunofluorescence staining for COL8A2 were performed. RESULTS: The cultured cells were immunofluorescently stained for collagen VIIIa1 and ZO-1. Collagenase-treated cultured human corneal endothelial cells formed monolayers on day 7 after transplantation into the rabbit corneal stroma and immunofluorescently stained for COL8A2 and ZO-1. The cultured human corneal endothelial cell sheet-rabbit stroma complex transplanted into rabbits was transparent on days 3 and 5, but corneal opacity developed by day 7. Histologic examination revealed 3,3′-dioctadecyloxacarbocyanine perchlorate (DIO)-stained corneal endothelium (green) and hard-tissue lymphocytes had infiltrated the cultured corneal endothelial cell plate-rabbit corneal stromal complex graft group. CONCLUSIONS: The cultured corneal endothelial cell sheet-rabbit corneal stromal complex prepared with the aid of collagenase showed a potential method for corneal endothelial cell transplantation.
Key Words: Collagenase;Corneal endothelial cell;Corneal transplantation;Cultured human corneal endothelial cell sheet;Endothelial cell transplantation


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