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羊膜作为培养的人角膜内皮细胞移植的载体

Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation.

作者信息

Ishino Yutaka, Sano Yoichiro, Nakamura Takahiro, Connon Che J, Rigby Helen, Fullwood Nigel J, Kinoshita Shigeru

机构信息

Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.

出版信息

Invest Ophthalmol Vis Sci. 2004 Mar;45(3):800-6. doi: 10.1167/iovs.03-0016.

Abstract

PURPOSE

It would be advantageous if cultivated human corneal endothelial cells (cHCECs) could be transplanted for the treatment of diseases caused by corneal endothelial disorders. To achieve this, a matrix that can serve as a carrier for cHCECs is needed. The present study was conducted to examine the feasibility of using amniotic membrane (AM) as a carrier for this application.

METHODS

HCECs obtained from peripheral corneal tissue were cultivated, passaged, and transplanted onto denuded AM. The cell density and morphology of the resultant cHCECs on AM were examined by light, scanning electron, and transmission electron microscopy. To determine whether these cHCEC sheets on AM carrier were functional in vivo, the cHCEC sheets on AM were transplanted onto rabbit corneas whose Descemet's membrane and endothelial cells had been completely removed. After transplantation, the corneal appearance was examined by slit lamp biomicroscopy, and corneal thickness was measured daily by pachymetry. At 7 days after surgery, the grafts were examined by light, scanning electron, and transmission electron microscopy.

RESULTS

The density of the cHCECs on AM was greater than 3000 cells/mm(2). Morphologically, the cHCEC sheets consisted of a fairly continuous layer of flat squamous polygonal endothelial cells that appeared uniform in size with tightly opposed cell junctions in vitro and in vivo after transplantation. The corneas that received transplanted cHCEC sheets had little edema and retained their thinness and transparency.

CONCLUSIONS

The cell density and morphology of cHCECs on AM were similar to those of normal corneas, and cHCECs on AM were functional in vivo. These results indicate that AM maintains HCEC morphology and function and could serve as a carrier for cHCEC transplantation.

摘要

目的

如果能够移植培养的人角膜内皮细胞(cHCECs)来治疗由角膜内皮疾病引起的病症将是有益的。要实现这一点,需要一种能够作为cHCECs载体的基质。本研究旨在检验使用羊膜(AM)作为该应用载体的可行性。

方法

从周边角膜组织获取的HCECs进行培养、传代,并移植到无细胞的AM上。通过光学显微镜、扫描电子显微镜和透射电子显微镜检查AM上所得cHCECs的细胞密度和形态。为了确定AM载体上的这些cHCEC片层在体内是否具有功能,将AM上的cHCEC片层移植到Descemet膜和内皮细胞已被完全去除的兔角膜上。移植后,通过裂隙灯生物显微镜检查角膜外观,并每天用测厚仪测量角膜厚度。术后7天,通过光学显微镜、扫描电子显微镜和透射电子显微镜检查移植物。

结果

AM上cHCECs的密度大于3000个细胞/mm²。在形态学上,cHCEC片层由相当连续的一层扁平鳞状多边形内皮细胞组成,这些细胞在体外和移植后的体内大小看起来均匀,细胞连接紧密相对。接受移植cHCEC片层的角膜几乎没有水肿,保持了其薄度和透明度。

结论

AM上cHCECs的细胞密度和形态与正常角膜相似,且AM上的cHCECs在体内具有功能。这些结果表明,AM维持了HCEC的形态和功能,并且可以作为cHCEC移植的载体。

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