Characterization of iris pigment epithelial cell for auto cell transplantation
Characterization of iris pigment epithelial cell for auto cell transplantation
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DOI：
10.1177/096368979900800505
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发表时间：
1999-09-01
影响因子：
3.3
通讯作者：
Tamai, M
中科院分区：
文献类型：
--
作者：
Abe, T;Tomita, H;Tamai, M
为建立自体虹膜色素上皮（IPE）细胞移植模型，我们对IPE细胞的特性及用自体血清培养IPE细胞的方法进行了研究。获得猴和人IPE细胞，并使用自体、小鼠、兔、牛或人血清在几种条件下培养。进行免疫细胞化学研究以确认细胞来源于上皮。IPE的增殖率也计算从新鲜的人IPE细胞，这是在滤过性青光眼手术中获得的。还将增殖率与视网膜色素上皮（RPE）细胞的增殖率进行比较。逆转录酶和聚合酶链反应的黑素生成进行，并在IPE细胞中的色素的量也进行了计算。小鼠和兔血清对猴IPE细胞培养无效。相反，细胞在含有自体血清、牛血清或人血清的培养基中生长良好。人IPE细胞通过所述方法呈指数生长，并在约4-5周后达到60，000个细胞。当我们比较它们的增殖率时，IPE细胞比RPE细胞的增殖率低。黑素生成的基因表达和IPE中的色素量通过连续传代逐渐下降。已尝试使用来自胎儿或眼库眼睛的RPE进行移植以治疗年龄相关性黄斑变性。然而，局灶性排斥反应可能在临床结果中起重要作用。自体IPE细胞移植的建立可能改善排斥反应问题。本研究利用自体血清建立了自体IPE细胞培养体系。培养的IPE细胞显示色素上皮细胞的特性，直到在人类和猴子中的约5代。
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 epiahelial (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.