State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Invest Ophthalmol Vis Sci. 2010 Nov;51(11):5970-8. doi: 10.1167/iovs.09-4504. Epub 2010 May 19.
Somatic cells can be reprogrammed into an embryonic stem cell-like pluripotent state by Oct-3/4, Sox2, c-Myc, and Klf4. Sox2 as an essential reprogramming factor also contributes to the development of the eye and the retina. This study was conducted to determine whether induced pluripotent stem (iPS) cells express retinal progenitor cell (RPC)-related genes and whether iPS cells can directly differentiate into retinal ganglion cells (RGCs).
Mouse iPS cells were induced by the ectopically expressed four factors in tail-tip fibroblasts (TTFs). The expression of RPC-related genes in iPS cells was analyzed by RT-PCR and immunofluorescence. iPS cells were induced to differentiate into RGCs by the addition of Dkk1 + Noggin (DN) + DAPT and overexpression of Math5. iPS-derived retinal ganglion (RG)-like cells were injected into the retina, and the eyes were analyzed by immunohistochemistry.
iPS cells inherently express RPC-related genes such as Pax6, Rx, Otx2, Lhx2, and Nestin. Overexpression of Math5 and addition of DN can directly differentiate iPS into retinal ganglion-like cells. These iPS-derived RG-like cells display long synapses and gene expression patterns, including Math5, Brn3b, Islet-1, and Thy1.2. Furthermore, inhibiting Hes1 by DAPT increases the expression of RGC marker genes. In addition, iPS-derived RG-like cells were able to survive but were unable to be integrated into the normal retina after transplantation.
The four factor iPS cell inherently expressed RPC-related genes, and the iPS cell could be further turned into RG-like cells by the regulation of transcription factor expression. These findings demonstrate that iPS cells are valuable for regeneration research into retinal degeneration diseases.
通过 Oct-3/4、Sox2、c-Myc 和 Klf4，体细胞可被重编程为胚胎干细胞样多能状态。Sox2 作为一种必需的重编程因子，也有助于眼睛和视网膜的发育。本研究旨在确定诱导多能干细胞（iPS）是否表达视网膜祖细胞（RPC）相关基因，以及 iPS 细胞是否能直接分化为视网膜神经节细胞（RGC）。
通过在尾尖成纤维细胞（TTFs）中外源表达这四种因子，诱导产生小鼠 iPS 细胞。通过 RT-PCR 和免疫荧光分析 iPS 细胞中 RPC 相关基因的表达。通过添加 Dkk1+Noggin（DN）+DAPT 和过表达 Math5 诱导 iPS 细胞分化为 RGC。将 iPS 来源的视网膜神经节（RG）样细胞注射到视网膜中，并通过免疫组织化学分析眼睛。
iPS 细胞固有地表达 RPC 相关基因，如 Pax6、Rx、Otx2、Lhx2 和 Nestin。过表达 Math5 和添加 DN 可直接将 iPS 分化为视网膜神经节样细胞。这些 iPS 衍生的 RG 样细胞显示出长突触和基因表达模式，包括 Math5、Brn3b、Islet-1 和 Thy1.2。此外，通过 DAPT 抑制 Hes1 可增加 RGC 标记基因的表达。此外，iPS 衍生的 RG 样细胞能够存活，但在移植后无法整合到正常的视网膜中。
四种因子 iPS 细胞固有地表达 RPC 相关基因，并且通过转录因子表达的调节，iPS 细胞可进一步转化为 RG 样细胞。这些发现表明 iPS 细胞在视网膜变性疾病的再生研究中具有重要价值。
