Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Kobe, Japan.
Invest Ophthalmol Vis Sci. 2011 Nov 11;52(12):8785-90. doi: 10.1167/iovs.11-8129.
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.
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.
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.
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.
诱导多能干细胞（iPS 细胞）有望成为再生医学研究与开发的有力工具。作者先前报道过，人 iPS 细胞可分化为视网膜细胞，包括光感受器和视网膜色素上皮细胞。在这项研究中，他们从猴子中产生 iPS 细胞系，以研究其分化为视网膜细胞的能力。
为了生成 iPS 细胞，作者将源自食蟹猴腹部皮肤的成纤维细胞用携带 Oct3/4、Sox2、Klf4 和 c-Myc 基因的逆转录病毒感染，然后在 STO 饲养细胞上培养。接着，用 PA6 细胞的条件培养基培养建立的 iPS 细胞以诱导 RPE 细胞。分析分化的 RPE 细胞的特性。
在病毒感染后约 1 个月，成纤维细胞中出现了一些上皮样集落。这些集落在形态上类似于食蟹猴胚胎干细胞（ES 细胞），并表达 ES 细胞特异性标志物。通过在 SCID 小鼠中产生畸胎瘤，证实这些细胞具有分化为三个胚层的能力。此外，从猴 iPS 细胞诱导而来的 RPE 细胞具有特征性的多角形形状和色素。这些细胞表达 RPE 细胞特异性标志物，如 RPE65、CRALBP、Bestrophin 1 和 MERTK，并在体外表现出吞噬功能。
利用 iPS 细胞技术从猴皮肤中获得的 RPE 细胞可用于自体或同种异体移植，以测试免疫排斥的可能性，并使用将在临床试验中使用的相同技术在体内评估其功能。
