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Characterization of iris pigment epithelial cell for auto cell transplantation.

机译:虹膜色素上皮细胞的特征用于自体细胞移植。

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To establish auto iris pigment epithelial (IPE) transplantation, we characterized the properties of IPE cells and the method of culture using auto serum. Monkey and human IPE cells were obtained and cultured in several conditions, using auto, mouse, rabbit, bovine, or human serum. Immunocytochemical study was performed to confirm that the cells were epithelial in origin. The proliferation rate of the IPE was also calculated from fresh human IPE cells, which were obtained during filtering glaucoma surgery. Proliferation rate was also compared to that of retinal pigment epithelial (RPE) cells. Reverse-transcriptase and polymerase chain reaction for melanogenesis was performed, and the amount of pigment in the IPE cells was also calculated. Mouse and rabbit sera were not effective for the monkey IPE cell culture. Conversely, the cells grew well in the medium with auto, bovine, or human serum. Human IPE cells grew exponentially by the described methods and reached to 60,000 cells after about 4-5 weeks. When we compared them by proliferation rate, IPE cells were less proliferative than RPE cells. The gene expression for melanogenesis and the amount of pigment in the IPE gradually decreased through successive passages. Transplantation has been tried for the treatment of age-related macular degeneration using RPE from fetus or from eye bank eyes. However, focal rejection may play an important role in the clinical results. The establishment of auto IPE cell transplantation may improve the problem of rejection. In the present study, we established auto IPE cell culture using auto serum. The cultured IPE cell showed pigment epithelial cell properties until around five passages in both human and monkey.
机译:为了建立自动虹膜色素上皮(IPE)移植,我们表征了IPE细胞的特性和使用自动血清的培养方法。获得猴和人IPE细胞,并在几种条件下使用汽车,小鼠,兔,牛或人血清培养。进行了免疫细胞化学研究,以确认细胞起源于上皮细胞。还从在过滤青光眼手术中获得的新鲜人IPE细胞计算IPE的增殖率。还将增殖速率与视网膜色素上皮(RPE)细胞的增殖速率进行了比较。进行了用于黑色素生成的逆转录酶和聚合酶链反应,并且还计算了IPE细胞中色素的量。小鼠和兔子的血清对猴子IPE细胞培养无效。相反,细胞在含有自身,牛或人血清的培养基中生长良好。通过描述的方法,人IPE细胞呈指数增长,并在大约4-5周后达到60,000个细胞。当我们通过增殖率比较它们时,IPE细胞的增殖性低于RPE细胞。通过连续传代,黑色素生成的基因表达和IPE中的色素含量逐渐降低。已经尝试使用来自胎儿或眼库眼睛的RPE进行移植治疗年龄相关性黄斑变性。但是,局灶性排斥反应可能在临床结果中起重要作用。自动IPE细胞移植的建立可以改善排斥反应的问题。在本研究中,我们使用自动血清建立了自动IPE细胞培养。培养的IPE细胞在人和猴子中直到大约五次传代都显示出色素上皮细胞特性。

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