Induction of Retinal Pigment Epithelial Cells from Monkey iPS Cells
Induction of Retinal Pigment Epithelial Cells from Monkey iPS Cells
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DOI：
10.1167/iovs.11-8129
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发表时间：
2011-11-01
影响因子：
4.4
通讯作者：
Takahashi, Masayo
中科院分区：
文献类型：
--
作者：
Okamoto, Satoshi;Takahashi, Masayo
目的。诱导多能干细胞(IPS)有望成为再生医学研究和开发的有力工具。此前，作者报道了人类iPS细胞分化为视网膜细胞，包括光感受器和视网膜色素上皮细胞。方法用携带Oct3/4、Sox2、Klf4和c-Myc基因的逆转录病毒感染食蟹猴腹部皮肤成纤维细胞，培养于STO饲养层细胞。接下来，将建立的iPS细胞与PA6细胞的条件培养液一起培养，以诱导RPE细胞。结果：病毒感染后约1个月，成纤维细胞间出现上皮样集落。这些克隆在形态上与食蟹胚胎干细胞相似，并表达ES细胞特异性标记。通过在SCID小鼠体内产生畸胎瘤，这些细胞被证实具有分化为三个胚层的能力。此外，从猴iPS细胞诱导的RPE细胞具有独特的多边形形状和色素。这些细胞表达RPE细胞特异性标志物RPE65、CRALBP、Bestrophin 1和MERTK，并在体外具有吞噬功能。结论采用iPS细胞技术从猴皮肤分离的RPE细胞可用于自体或同种异体移植，以检测免疫排斥反应的可能性并评估其体内功能，该技术将用于临床试验。(Invest Ophthalol Vis Science.2011；52：8785-8790)doi：10.1167/iovs.11-8129
PURPOSE. The induced pluripotent stem (iPS) cell is expected to be a powerful tool for research and development in regenerative medicine. Previously, the authors reported that human iPS cells differentiated into retinal cells, including photoreceptors and retinal pigment epithelial cells. In this study, they produced iPS cell lines from monkeys to investigate their ability to differentiate into retinal cells.METHODS. To generate iPS cells, the fibroblasts derived from cynomolgus monkey abdominal skin were infected with retroviruses carrying Oct3/4, Sox2, Klf4, and c-Myc genes and then were cultured on STO feeder cells. Next, the established iPS cells were cultured with the conditioned medium of PA6 cells to induce RPE cells. The properties of the differentiated RPE cells were analyzed.RESULTS. Approximately 1 month after viral infection, some epithelial-like colonies appeared among the fibroblasts. These colonies were morphologically similar to the cynomolgus embryonic stem (ES) cell and expressed ES cell-specific markers. By producing teratomas in SCID mice, these cells were confirmed to have the ability to differentiate into three germ layers. In addition, the RPE cells induced from the monkey iPS cells had characteristic polygonal shapes and pigments. These cells expressed RPE cell-specific markers such as RPE65, CRALBP, Bestrophin 1, and MERTK and exhibited phagocytotic function in vitro.CONCLUSIONS. The RPE cells derived from monkey skin with iPS cell technology can be used for autologous or allogeneic transplantation to test the possibility of immune rejection and to evaluate their function in vivo with the same techniques that will be used in clinical trials. (Invest Ophthalmol Vis Sci.2011;52:8785-8790) DOI:10.1167/iovs.11-8129