Department of Regenerative Medicine, Graduate School of Medicine, University of the Ryukyus, 207 Uehara, Nishihara, Okinawa 903-0215, Japan.
Department of Surgery, Okayama Saidaiji Hospital, Okayama 704-8192, Japan.
Int J Mol Sci. 2018 Nov 6;19(11):3489. doi: 10.3390/ijms19113489.
Adipose-derived mesenchymal stem cells (ADSCs) have attracted attention due to their potential for use in the treatment of various diseases. However, the self-renewal capacity of ADSCs is restricted and their function diminishes during passage. We previously generated induced tissue-specific stem cells from mouse pancreatic cells using a single synthetic self-replicating Venezuelan Equine Encephalitis (VEE)-reprogramming factor (RF) RNA replicon (SR-RNA) expressing the reprogramming factors POU class 5 homeobox 1 (OCT4), Krueppel-like factor 4 (KLF4), Sex determining region Y-box 2 (SOX2), and Glis Family Zinc Finger 1 (GLIS1). This vector was used to generate induced pluripotent stem (iPS) cells. Here, we applied this SR-RNA vector to generate human iTS cells from aged mesenchymal stem cells (hiTS-M cells) deficient in self-renewal that were derived from adipose tissue. These hiTS-M cells transfected with the SR-RNA vector survived for 15 passages. The hiTS-M cells expressed cell surface markers similar to those of human adipose-derived mesenchymal stem cells (hADSCs) and differentiated into fat cells and osteoblasts. Global gene expression profiling showed that hiTS-M cells were transcriptionally similar to hADSCs. These data suggest that the generation of iTS cells has important implications for the clinical application of autologous stem cell transplantation.
脂肪来源间充质干细胞（ADSCs）因其在治疗各种疾病方面的应用潜力而备受关注。然而，ADSCs 的自我更新能力有限，在传代过程中其功能会逐渐减弱。我们之前曾使用一种表达重编程因子 POU 类 5 同源框 1（OCT4）、Krüppel 样因子 4（KLF4）、性决定区 Y 盒 2（SOX2）和 Glis 家族锌指 1（GLIS1）的单种合成组织特异性干细胞的委内瑞拉马脑炎（VEE）复制型 RNA 复制子（SR-RNA）从胰腺细胞中诱导产生组织特异性干细胞。该载体用于生成诱导多能干细胞（iPS 细胞）。在这里，我们将这种 SR-RNA 载体应用于从脂肪组织中分离的自我更新能力缺陷的衰老间充质干细胞（hiTS-M 细胞）中生成人诱导组织特异性干细胞（hiTS-M 细胞）。这些用 SR-RNA 载体转染的 hiTS-M 细胞可存活 15 代。hiTS-M 细胞表达与人类脂肪来源间充质干细胞（hADSCs）相似的细胞表面标志物，并分化为脂肪细胞和成骨细胞。全基因表达谱分析表明，hiTS-M 细胞在转录水平上与 hADSCs 相似。这些数据表明，iTS 细胞的生成对自体干细胞移植的临床应用具有重要意义。
