Cultured human corneal endothelial cell transplantation with a collagen sheet in a rabbit model
Cultured human corneal endothelial cell transplantation with a collagen sheet in a rabbit model
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DOI：
10.1167/iovs.03-1174
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发表时间：
2004-09-01
影响因子：
4.4
通讯作者：
Amano, S
中科院分区：
文献类型：
--
作者：
Mimura, T;Yamagami, S;Amano, S
目的。目的探讨体外培养的人角膜内皮细胞(HCECs)的体内功能及以胶原片作为HCEC替代载体移植的可行性。方法：用DiI(1，1‘-二十八烷基-3，3，3’，3‘-四甲基吲哚碳菁高氯酸盐)标记培养的成人供体角膜，接种于胶原片上。通过测量电位差和短路电流来评价HCEC片材的泵浦功能。在兔眼上进行6 mm硬化性角膜切口术和后角膜切开术。将胶原片上的HCEC带入前房并固定于后基质(HCEC组)。对照组为兔角膜片移植组(胶原组)和单纯角膜片移植组(非移植组)。每组术后观察28天，进行组织学和荧光显微镜检查。结果：HCEC片泵功能参数是人类供体角膜的76%~95%。术后第1、3、7、14、21、28天，HCEC组平均角膜厚度明显低于胶原组和未移植组(P<；0.05)。在HCEC组，DiI标记细胞散布在角膜后表面。苏木精-伊红染色结果显示，胶原组和未移植组均可见明显的基质水肿，而含有单层细胞胶原片的HCEC组则未见明显的基质水肿。结论本研究结果表明，从成人供体角膜移植的培养的HCECs在兔模型中仍能保留其角膜脱水的功能，提示用含胶原片的培养的HCEC移植治疗CEC功能障碍是可行的。
PURPOSE. To evaluate the function of cultured human corneal endothelial cells (HCECs) in vivo and the feasibility of HCEC transplantation with a collagen sheet as the substitute carrier of HCECs.METHODS. Adult human donor cornea derived from cultured HCECs was labeled with the fluorescent tracker DiI (1,1'-dioctadecyl- 3,3,3',3'-tetramethylindocarbocyanine perchlorate) and seeded on a collagen sheet. The pump function of the HCEC sheet was evaluated by measurement of the potential difference and short-circuit current. A 6-mm sclerocorneal incision and Descemetorhexis were performed on rabbit eyes. The HCECs on a collagen sheet was brought into the anterior chamber and fixed to the posterior stroma (HCEC group). Rabbit corneas with collagen sheet transplantation after Descemetorhexis (collagen group) and with only Descemetorhexis (no-transplantation group) were the control. Each group, observed for 28 days after surgery, underwent histologic and fluorescence microscopic examinations.RESULTS. Pump function parameters of the HCEC sheets were 76% to 95% of those of human donor corneas. Mean corneal thickness in the HCEC group was significantly less than in the collagen and no-transplantation groups 1, 3, 7, 14, 21, and 28 days (P < 0.05) after surgery. DiI-labeled cells were spread over the rear corneal surface in the HCEC group. Marked stromal edema was present in the collagen and no-transplantation groups with hematoxylin-eosin staining, but none in the HCEC group with collagen sheets bearing monolayer cells.CONCLUSIONS. The findings indicate that cultured HCECs transplanted from adult human donor cornea by means of a collagen sheet can retain their function of corneal dehydration in a rabbit model and suggest the feasibility of transplantation for CEC dysfunction using cultured HCECs with a collagen sheet.