人脐静脉血管内皮细胞在体移植替代猴角膜内皮细胞的形态学分析  

Microstructure observation of human vascular endothelial cells transplantation in autograft to replace monkey corneal endothelial cells

作　　者：张文佳[1] 胡竹林[1] 朱勤[1] 韦春玲[1] 孙晓梅[2] 代解杰[2] 刘海[1] 杨忠昆[1] 

机构地区：[1]昆明医科大学第四附属医院眼科,云南省昆明市650021 [2]中国医学科学院中国协和医科大学医学生物学研究所,云南省昆明市650000

出　　处：《眼科新进展》2014年第11期1020-1024,共5页Recent Advances in Ophthalmology

基　　金：2013年云南省科技厅昆明医科大学应用基础研究联合专项(编号:2013FB109)~~

摘　　要：目的使用去除后弹力层的恒河猴自体角膜为细胞载体,将培养的人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVEC)种植到角膜内表面,观察HUVEC替代恒河猴角膜内皮细胞的功能情况以及HUVEC在恒河猴眼内生长的情况。方法取恒河猴6只随机分为3组:实验组(3只)、实验对照组(2只)、空白对照组(1只)。实验组:用离心沉淀法将培养的HUVEC移植到去除后弹力层的恒河猴自体角膜内表面,之后将自体角膜缝回植床;实验对照组:将撕除部分后弹力层的术眼角膜植片原位缝回植床;空白对照组:取下术眼角膜植片不做任何处理原位缝回植床。术后观察各组角膜植片透明情况;实验组及实验对照组于术后30 d及60 d、空白对照组于术后60 d行术眼摘除,标本做病理切片、CD34免疫组化及扫描电镜,观察房角结构及HUVEC在角膜植片内表面形态分布。结果实验组角膜植片维持了一定的厚度和透明性,而实验对照组角膜植片发生严重大泡性改变。病理切片示实验组角膜内表面可见一层细胞生长,CD34染色阳性,提示为血管内皮细胞;实验对照组角膜内表面未见任何细胞生长;空白对照组角膜植片保留完整后弹力层及内皮细胞层。扫描电镜示实验组有HUVEC单层在角膜内表面生长但有大量白细胞聚集及少量细胞碎片嵌顿于小梁网;实验对照组角膜内表面残留胶原纤维样物质,无细胞生长;空白对照组见完整六边形角膜内皮细胞层。结论 HUVEC能够在撕除后弹力层的恒河猴角膜内表面生长并发挥一定的屏障作用,维持角膜的厚度和透明性,但会产生较重的排斥反应。Objective Using the Descemet＇s membrane-removed rhesus autologous corneal as the cell carrier and transplant the cultured human umbilical vein endothelial cells（ HUVEC） to the inner surface of the cornea,to observe the function of HUVEC after alternative rhesus monkey corneal endothelial cell and the growth of the HUVEC in the rhesus monkey eye corneal surface.Methods The experimented monkeys were divided into 3 groups： experimental group（ 3 cases）,experimental control group（ 2 cases） and blank control group（ 1 case）. In experimental group,cultured HUVEC were transplanted onto rhesus monkey posterior corneal surface without corneal endothelium; In experimental control group,the detached corneal Descemet membrane of rhesus monkey fixed in suit without cultured HUVEC transplantation; In blank control group,a simple penetrativity corneal transplantation without destroy the descemet membrane performed. The transparent of implants in each group was observed after corneal transplantation. Histopathology of cornea grafts,immunohistochemistry and scanning electron microscope were performed at 30 days and 60 days postoperatively in order to observe the appearance of HUVEC onto the monkey posterior corneal surface and the structure of anterior chamber angle. Results The corneal implants of experimental group kept transparency and thickness in some extent while the experimental control group showed severity corneal edema and bullae. Histopathology of corneal grafts showed that a cells layer with CD34 staining positive reaction growth on posterior corneal surface of corneal button in the experimental group,which indicated that the layer of cells were HUVEC,no cell and material like membrane could be seen on that of the experimental control group; A layer of corneal endothelial cells and Descemet＇s membrane were visible on that of the blank control group.Scanning electron microscope showed that a layer of HUVEC were growing on the posterior surface of cornea button with plenty of leucocytes and small amo

关 键 词：血管内皮细胞 角膜内皮细胞 恒河猴 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]