Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.
J Proteome Res. 2011 Mar 4;10(3):977-89. doi: 10.1021/pr100624f. Epub 2011 Feb 4.
The recent generation of induced pluripotent stem (iPS) cells represents a novel opportunity to complement embryonic stem (ES) cell-based approaches. iPS cells can be generated by viral transduction of specific transcription factors, but there is a potential risk of tumorigenicity by random retroviral integration. We have generated novel iPS (sFB-protein-iPS) cells from murine dermal fibroblasts (FVB-sFB) that have ES cell characteristics, using ES cell-derived cell extracts instead of performing viral transduction. Notably, only cell extracts from an ES cell line (C57-mES) on the C57/BL6 background generated iPS cells in our protocol-not an ES cell line (E14-mES) on the 129 background. Hypothesizing that determining the differences in these 2 mES cell lines will provide vital insight into the reprogramming machinery, we performed proteomic and global gene expression analysis by iTRAQ and mRNA microarray, respectively. We observed that pluripotent ES cells and ES cell extract-derived iPS cells had differential proteomes and global gene expression patterns. Notably, reprogramming-competent C57-mES cells highly expressed proteins that regulate protein synthesis and metabolism, compared with reprogramming-incompetent 129-mES cells, suggesting that there is a threshold that protein synthetic machinery must exceed to initiate reprogramming.
最近一代的诱导多能干细胞（iPS）代表了一种补充胚胎干细胞（ES）细胞方法的新机会。iPS 细胞可以通过特定转录因子的病毒转导产生，但随机逆转录病毒整合存在潜在的致瘤风险。我们使用 ES 细胞衍生的细胞提取物而不是进行病毒转导，从 FVB 来源的皮肤成纤维细胞（FVB-sFB）中生成了具有 ES 细胞特征的新型 iPS（sFB-protein-iPS）细胞。值得注意的是，只有来自 C57/BL6 背景的 ES 细胞系（C57-mES）的细胞提取物在我们的方案中生成 iPS 细胞，而不是来自 129 背景的 ES 细胞系（E14-mES）。假设确定这两个 mES 细胞系之间的差异将为重编程机制提供重要的见解，我们分别通过 iTRAQ 和 mRNA 微阵列进行了蛋白质组学和全基因表达分析。我们观察到多能 ES 细胞和 ES 细胞提取物衍生的 iPS 细胞具有不同的蛋白质组和全基因表达模式。值得注意的是，与重编程无能的 129-mES 细胞相比，具有重编程能力的 C57-mES 细胞高度表达调节蛋白质合成和代谢的蛋白质，这表明存在一个阈值，蛋白质合成机制必须超过该阈值才能启动重编程。
