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

发布时间：2018-04-28 11:46

本文选题：血管内皮细胞 + 角膜内皮细胞　；参考：《眼科新进展》2014年11期

【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。
[Abstract]:Objective to implant cultured human umbilical vein endothelial cells (HUVECs) of human umbilical vein endothelial cells (HUVECs) onto the inner surface of cornea using rhesus monkey autologous cornea after removing the elastic layer. The function of corneal endothelial cells replaced by HUVEC and the growth of HUVEC in the eyes of rhesus monkeys were observed. Methods six rhesus monkeys were randomly divided into three groups: experimental group (n = 3), experimental control group (n = 2) and blank control group (n = 1). In the experimental group, the cultured HUVEC was transplanted to the inner surface of the autogenous cornea of the rhesus monkey with the removal of the posterior elastic layer by centrifugal precipitation method, and then the corneal autograft was sewn back into the bed, while in the experimental control group, the corneal graft was sewn back into the bed in situ. Blank control group: removal of corneal graft without any treatment in situ sewn back to the bed. The corneal grafts in the experimental group and the experimental control group were observed on the 30th and 60th day after operation, and the blank control group were excised on the 60th day after operation, and the specimens were stained with CD34 immunohistochemistry and scanning electron microscope (SEM). The angle structure and the distribution of HUVEC on the surface of cornea grafts were observed. Results the corneal grafts in the experimental group maintained a certain thickness and transparency, while the corneal grafts in the experimental control group showed severe vesicular changes. Pathological sections showed that a layer of cells was positive for CD34 staining on the inner surface of cornea in the experimental group and no cell growth was found on the inner surface of the cornea in the experimental control group. The intact posterior elastic layer and endothelial cell layer were retained in the control group. Scanning electron microscope (SEM) showed that HUVEC monolayer grew on the inner surface of cornea but a large number of white blood cells were accumulated and a small amount of cell fragments were embedded in trabecular meshwork, while in the control group, collagen fiber like substance remained on the inner surface of cornea without cell growth. The intact hexagonal corneal endothelial cell layer was found in the blank control group. Conclusion HUVEC can grow on the inner surface of cornea of rhesus monkey after tearing out elastic layer and play a certain role as barrier to maintain the thickness and transparency of cornea, but it will produce serious rejection.
【作者单位】：昆明医科大学第四附属医院眼科;中国医学科学院中国协和医科大学医学生物学研究所;
【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)~~
【分类号】：R772.21

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