| 目的：探讨以人羊膜基底膜为细胞载体培养角膜内皮细胞，及进行角膜内皮细胞移植的可行性。
方法：首先以加工处理的人羊膜基底膜为细胞载体，种植培养猪眼角膜内皮细胞；取传3~5代的角膜内皮细胞单层，行猫眼角膜内皮细胞移植；术后不同时间应用光电镜观察植入的内皮细胞单层及羊膜载体的形态变化及功能。
结果：（1）倒置显微镜及扫描电镜观察：培养的猪眼角膜内皮细胞能在人羊膜上形成一完整的内皮细胞单层，细胞密度为2200个/mm2 ；（2）裂隙灯显微镜观察人羊膜为载体培养的角膜内皮细胞植入去除自体角膜内皮细胞的猫眼，术后6~12h植片水肿开始减轻，透明度增加；24~48h后植片恢复透明,维持正常角膜厚度;3周内植片保持透明无水肿；（3）术后不同时间取植片进行光、电镜观察表明，植入的内皮细胞可能在猫活体内发挥正常活体角膜内皮细胞的“液泵”功能；（4）术后3~4周均出现明显免疫排斥反应而致植片水肿、混浊。
结论：以人羊膜为载体行培养的单层内皮细胞移植术后，植入内皮能在活体前房环境中成活,保持正常形态并发挥与正常角膜内皮细胞相似的生理功能，为临床角膜移植开辟了一条新的途径。
关键词：角膜内皮细胞移植；羊膜；细胞培养；猫
Transplatation of Corneal Endothelium Cultured on the basic fundus of processed human aminiotic membrane
Purpose: To investigate the feasibility of transplantation of the cultured porcine corneal endothelial cells using a cell carrier device-processed human aminiotic membrane.
Methods: At first, the cultured porcine corneal endothelial cells were cultured on the processed human aminiotic membrane until they had reached a complete monolayer with an average of cell density of 2200±250 cells/mm2; and then ,the correlated cultured endothelia were transplanted into cats to replace the feline native corneal Descement’s membrane and endothelium via penetrating keratoplasty. Finally, the morphology and physiological function of the transplanted corneal endothelium were clinically evaluated by means of light and electron microscopic examination.
Results: Light and scanning electron microscopy showed that the cultured porcine corneal endothelial cells could in vitro form a complete endothelial monolayer on the carrier. After operation, slim microscope examination showed that all grafts recovered clean and remained clear for an average of three weeks without signs of rejection or inflammation in vivo. Light and electron microscopic examination have demonstrated that the transplanted endothelial cells presented regular hexagonal pattern. And however, 3-4 weeks postoperatively, the observed grafts appear edematous, opaque and vascularized severly, which correlated with the light and electron microscopic findings .
Conclusion: The research demonstrated that corneal endothelial cells cultured in vitro on processed human aminiotic membrane could be replaced and remain functional adequately for at least three weeks, which suggested that this technique could be ultimately developed for routine use in human.
Key words: corneal endothelial cell transplantation; human amniotic membrane; cellular culture; cat.
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