Differential Effects of Extracellular Vesicles of Lineage-Specific Human Pluripotent Stem Cells on the Cellular Behaviors of Isogenic Cortical Spheroids
Differential Effects of Extracellular Vesicles of Lineage-Specific Human Pluripotent Stem Cells on the Cellular Behaviors of Isogenic Cortical Spheroids
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作者：
Marzano, Mark;Bejoy, Julie;Li, Yan
细胞外囊泡（EV）有助于多种信号传导过程以及干细胞和组织的整体生理和病理状态。人类诱导多能干细胞（hiPSC）具有能够模拟胚胎组织发育的独特特性。人们越来越关注将源自hiPSC的EV用作治疗剂、生物标志物和药物递送载体。然而，很少有人知道由hiPSC分泌的EV和旁分泌信号在组织形态发生和谱系特化的特征。研究方法：在这项研究中，分析了从hiPSC衍生的神经祖细胞（外胚层）、hiPSC衍生的心脏细胞（中胚层）和未分化的hiPSC（健康iPSK 3和阿尔茨海默病相关的SY-UBH系）分离的EV的物理和生物学性质。结果：纳米颗粒跟踪分析和电子显微镜结果表明hiPSC衍生的EV具有100-250 nm的平均尺寸。免疫印迹分析证实了纯化EV制剂中外来体标记物阿利克斯、CD 63、TSG 101和Hsc 70的富集。包括miR-133、miR-155、miR-221和miR-34 a的微RNA在分离自不同hiPSC谱系的EV中不同地表达。在体外用hiPSC-EV处理皮质球状体导致增强的细胞增殖（由BrdU+细胞指示）和轴突生长（由β-微管蛋白III染色指示）。此外，hiPSC衍生的EV在A β 42寡聚体处理的培养物中表现出神经保护能力，增强细胞活力并降低氧化应激。我们的研究结果表明，由来自hiPSC的组织上下文依赖性EV提供的旁分泌信号传导引起不同的反应，以影响皮质球状体的生理状态。总的来说，这项研究推进了我们对干细胞微环境中细胞间通讯的理解，并为治疗神经退行性变提供了可能的治疗选择。
Extracellular vesicles (EVs) contribute to a variety of signaling processes and the overall physiological and pathological states of stem cells and tissues. Human induced pluripotent stem cells (hiPSCs) have unique characteristics that can mimic embryonic tissue development. There is growing interest in the use of EVs derived from hiPSCs as therapeutics, biomarkers, and drug delivery vehicles. However, little is known about the characteristics of EVs secreted by hiPSCs and paracrine signaling during tissue morphogenesis and lineage specification. Methods: In this study, the physical and biological properties of EVs isolated from hiPSC-derived neural progenitors (ectoderm), hiPSC-derived cardiac cells (mesoderm), and the undifferentiated hiPSCs (healthy iPSK3 and Alzheimer's-associated SY-UBH lines) were analyzed. Results: Nanoparticle tracking analysis and electron microscopy results indicate that hiPSC-derived EVs have an average size of 100-250 nm. Immunoblot analyses confirmed the enrichment of exosomal markers Alix, CD63, TSG101, and Hsc70 in the purified EV preparations. MicroRNAs including miR-133, miR-155, miR-221, and miR-34a were differently expressed in the EVs isolated from distinct hiPSC lineages. Treatment of cortical spheroids with hiPSC-EVs in vitro resulted in enhanced cell proliferation (indicated by BrdU+ cells) and axonal growth (indicated by beta-tubulin III staining). Furthermore, hiPSC-derived EVs exhibited neural protective abilities in A beta 42 oligomer-treated cultures, enhancing cell viability and reducing oxidative stress. Our results demonstrate that the paracrine signaling provided by tissue context-dependent EVs derived from hiPSCs elicit distinct responses to impact the physiological state of cortical spheroids. Overall, this study advances our understanding of cellcell communication in the stem cell microenvironment and provides possible therapeutic options for treating neural degeneration.