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Reconstruction of corneal epithelium with cryopreserved corneal limbal stem cells in a rabbit model
Authors:Qu Lei  Yang Xueyi  Wang Xin  Zhao Ming  Mi Shengli  Dou Zhongying  Wang Huayan
Institution:aCollege of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi 712100, China;bLife Science Research Centre, Yulin College, Yulin, Shaanxi 719000, China;cDepartment of Ophthalmology, Yangling Hospital, Yangling, Shaanxi 712100, China
Abstract:The integrity and transparency of the cornea plays a key role in preserving vision. This paper reports a procedure to create an artificial sheet of corneal epithelium from cryopreserved limbal stem cells (LSCs) and to use this for corneal transplantation. Corneal LSCs were isolated from biopsy specimens of rabbit limbal lamellar and cryopreserved in liquid nitrogen at 2–4 passages. The cells were grown in culture medium for 12–14 days on top of a cell-free human amniotic membrane framed on a nitrocellulose sheet. The corneal epithelium generated was transplanted into the right eyes of 14 LSC deficient (LSCD) rabbits (seven experimental animals, seven controls) with corneal damage. The seven LSCD rabbits in the experimental group were transplanted with a corneal epithelial sheet generated from the cryopreserved corneal LSCs. Four LSCD rabbits were used as the vehicle control and were transplanted with a cell-free amniotic membrane, and the remaining three LSCD rabbits were negative controls without transplantation. Over a 2-month recovery period, 2/7 animals in the experimental group recovered completely, four recovered partially and one did not respond. In the control groups, three negative controls and three vehicle controls lost their vision completely, and one of the vehicle controls partially recovered transparency of the cornea Following treatment, corneal transparency of the experimental rabbits was significantly improved compared to controls (P < 0.05). The results indicated that cryopreserved corneal LSCs can repair damaged rabbit cornea, suggesting a possible new clinical approach to reconstruction of corneal epithelium.
Keywords:Amniotic membrane  Cornea  Cryopreservation  Limbal stem cells  Rabbit
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