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Abstract：
Human induced pluripotent stem cells (hiPSCs) have been regarded as a potential stem cell source for cell therapy. However, the production of cells with mesenchymal potential from hiPSCs through spontaneous differentiation is time consuming and laborious. In the present study, the combined use of the GSK-3 inhibitor CHIR99021 and TGF-β was used to obtain mesenchymal stem cell (MSC)-like cells from hiPSCs. During the induction process, the transcription of epithelial-mesenchymal transition (EMT)-related genes N-cadherin and Vimentin in the transformed cells was upregulated, whereas the transcription of E-cadherin and pluripotency-related transcription factors SOX2, OCT4 and NANOG did not change significantly. This indicated that whilst cells were pluripotent, EMT was initiated by the upregulation of transcription of EMT promoting genes. Both SMAD-dependent and independent signalling pathways were significantly activated by the combined induction treatment compared with the single factor induction. The hiPSC-derived MSC-like cells (hiPSC-MSCs) expressed MSC-related markers and acquired osteogenic, chondrogenic and adipogenic differentiation potentials. After being injected into the peritoneal cavity of rats, the hiPSC-MSCs secreted angiogenic and immune-regulatory factors and remained on the colicomentum for 3 weeks. Within an 11-week period, four intraperitoneal hiPSC-MSC injections (1x107 cells/injection) into acute myocardial infarction (AMI) model rats significantly increased the left ventricular ejection fraction, left ventricular fractional shortening and angiogenesis and significantly reduced scar size and the extent of apoptosis in the infarcted area compared with that of the control PBS injection. Symptoms of hiPSC-MSC-induced immune reaction or tumour formation were not observed over the course of the experiment in the hiSPC-MSC treated rats. In conclusion, the CHIR99021 and TGF-β combined induction was a rapid and effective method to obtain MSC-like cells from hiPSCs and multiple high dose intraperitoneal injections of hiPSC-derived MSCs were safe and effective at restoring cardiac function in an AMI rat model.
摘要：
人诱导多能干细胞(hiPSC)已被认为是用于细胞治疗的潜在干细胞来源。然而,通过自发分化从hiPSC产生具有间充质潜能的细胞是耗时且费力的。在本研究中,GSK-3抑制剂CHIR99021和TGF-β的联合使用用于从hiPSC获得间充质干细胞（MSC）样细胞。在感应过程中,转化细胞中上皮-间质转化（EMT）相关基因N-cadherin和波形蛋白的转录上调，而E-cadherin和多能性相关转录因子SOX2,OCT4和NANOG的转录没有显著变化.这表明虽然细胞是多能的，EMT由EMT促进基因转录的上调启动。与单因素诱导相比，联合诱导治疗可显著激活SMAD依赖性和非依赖性信号通路。hiPSC来源的MSC样细胞(hiPSC-MSCs)表达MSC相关标志物,软骨形成和脂肪形成分化潜能。注射到大鼠的腹膜腔后，hiPSC-MSCs分泌血管生成和免疫调节因子,并在结肠网膜上停留3周.在11周内,4次腹腔注射hiPSC-MSC(1x107细胞/注射)急性心肌梗死(AMI)模型大鼠左心室射血分数,与对照PBS注射相比，左心室缩短分数和血管生成以及梗死区瘢痕大小和凋亡程度显着降低。在hiPSC-MSC治疗的大鼠中,在实验过程中未观察到hiPSC-MSC诱导的免疫反应或肿瘤形成的症状。总之,CHIR99021和TGF-β联合诱导是一种从hiPSCs中获得MSC样细胞的快速有效方法,在AMI大鼠模型中,腹膜内多次大剂量注射hiPSC来源的MSC对恢复心功能是安全有效的.
