Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation
Amniotic membrane as a carrier for cultivated human corneal endothelial cell transplantation
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DOI：
10.1167/iovs.03-0016
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发表时间：
2004-03-01
影响因子：
4.4
通讯作者：
Kinoshita, S
中科院分区：
文献类型：
--
作者：
Ishino, Y;Sano, Y;Kinoshita, S
目的。培养的人角膜内皮细胞（cHCECs）可用于角膜内皮疾病的移植治疗。为了实现这一目标，需要一种可以作为cHCECs载体的矩阵。以羊膜为载体，探讨其应用的可行性。从角膜周围组织中获得的HCECs被培养、传代并移植到去角质的AM上。通过光镜、扫描电镜和透射电镜观察AM上所得cHCECs的细胞密度和形态。为了确定AM载体上的cHCEC片在体内是否具有功能，我们将AM载体上的cHCEC片移植到完全去除Descemet膜和内皮细胞的兔角膜上。移植后用裂隙灯生物显微镜观察角膜外观，每日用角膜厚度仪测量角膜厚度。术后1天，行光镜、扫描电镜、透射电镜检查。AM上cHCECs密度大于3000个/mm(2)。形态学上，移植后的cHCEC片由相当连续的扁平鳞状多角形内皮细胞组成，细胞大小均匀，细胞连接紧密相反。移植cHCEC片的角膜水肿少，保持了角膜的薄度和透明度。AM上的cHCECs细胞密度和形态与正常角膜相似，在体内具有一定的功能。这些结果表明AM维持了HCEC的形态和功能，可以作为HCEC移植的载体。
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 1 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.