Zhang Yusen, Zhang Yanmin, Hu Azhen, Meng Fanhua, Cui Peng, Li Tianshi, Cui Guanghui
Department of Ultrasound, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China.
Central Laboratory, Peking University Shenzhen Hospital, Shenzhen, Guangdong 518036, P.R. China.
Exp Ther Med. 2024 Mar 4;27(5):182. doi: 10.3892/etm.2024.12470. eCollection 2024 May.
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 and 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 (1x10 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.
人诱导多能干细胞（hiPSC）被视为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既耗时又费力。在本研究中，联合使用GSK-3抑制剂CHIR99021和转化生长因子-β（TGF-β）从hiPSC获得间充质干细胞（MSC）样细胞。在诱导过程中，转化细胞中上皮-间充质转化（EMT）相关基因N-钙黏蛋白和波形蛋白的转录上调，而E-钙黏蛋白和多能性相关转录因子的转录没有显著变化。这表明虽然细胞具有多能性，但EMT是由EMT促进基因转录上调引发的。与单因素诱导相比，联合诱导处理显著激活了SMAD依赖和非依赖信号通路。hiPSC来源的MSC样细胞（hiPSC-MSC）表达MSC相关标志物，并获得了成骨、成软骨和脂肪生成分化潜能。将hiPSC-MSC注射到大鼠腹腔后，它们分泌血管生成和免疫调节因子，并在结肠系膜上停留3周。在11周内，与对照PBS注射相比，向急性心肌梗死（AMI）模型大鼠腹腔内注射四次hiPSC-MSC（每次注射1×10个细胞）显著提高了左心室射血分数、左心室缩短分数和血管生成，并显著减小了梗死面积和梗死区域的凋亡程度。在实验过程中，未观察到hiPSC-MSC处理的大鼠出现hiPSC-MSC诱导的免疫反应或肿瘤形成症状。总之，CHIR99021和TGF-β联合诱导是从hiPSC获得MSC样细胞的快速有效方法，多次高剂量腹腔注射hiPSC来源的MSC对恢复AMI大鼠模型的心脏功能是安全有效的。
