Extracellular vesicles (EVs) regulate cell metabolism and various biological processes by delivering specific proteins and nucleic acids to surrounding cells. We aimed to investigate the effects of the cargo contained in EVs derived from adipose-derived stem cells (ASCs) on the porcine embryonic development. ASCs were isolated from porcine adipose tissue and characterized using ASC-specific markers via flow cytometry. EVs were subsequently extracted from the conditioned media of the established ASCs. These EVs were added to the in vitro culture environment of porcine embryos to observe qualitative improvements in embryonic development. Furthermore, the proteins within the EVs were analyzed to investigate the underlying mechanisms. We observed a higher blastocyst development rate and increased mitochondrial activity in early stage embryos in the ASC-EVs-supplemented group than in the controls (24.8% ± 0.8% vs. 28.6% ± 1.1%, respectively). The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay of blastocysts also revealed significantly reduced apoptotic cells in the ASC-EVs-supplemented group. Furthermore, through proteomics, we detected the proteins in ASC-EVs and blastocysts from each treatment group. This analysis revealed a higher fraction of proteins in the ASC-EVs-supplemented group than in the controls (1,547 vs. 1,495, respectively). Gene analysis confirmed that ASC-EVs showed a high expression of tyrosine-protein kinase (SRC), whereas ASC-EVs supplemented blastocysts showed a higher expression of Cyclin-dependent kinase 1 (CDK1). SRC is postulated to activate protein kinase B (AKT), which inhibits the forkhead box O signaling pathway and activates CDK1. Subsequently, CDK1 activation influences the cell cycle, thereby affecting in vitro embryonic development. ASC-EVs promote mitochondrial activity, which is crucial for the early development of blastocysts and vital in the downregulation of apoptosis. Additionally, ASC-EVs supply SRC to porcine blastocysts, thereby elongating the cell cycle.

中文翻译：

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含脂肪的外源蛋白干细胞囊泡的胚胎营养作用


细胞外囊泡 （EV） 通过将特定蛋白质和核酸输送到周围细胞来调节细胞代谢和各种生物过程。我们旨在研究脂肪来源干细胞 （ASCs） 衍生的 EVs 中所含的货物对猪胚胎发育的影响。从猪脂肪组织中分离 ASCs，并通过流式细胞术使用 ASC 特异性标记物进行表征。随后从已建立的 ASC 的条件培养基中提取 EV。将这些 EV 添加到猪胚胎的体外培养环境中，以观察胚胎发育的质的改善。此外，还分析了 EV 内的蛋白质以研究潜在机制。我们观察到补充 ASC-EVs 组的早期胚胎囊胚发育速率和线粒体活性增加（分别为 24.8% ± 0.8% vs. 28.6% ± 1.1%）。囊胚的末端脱氧核苷酸转移酶 dUTP 缺口末端标记 （TUNEL） 测定也显示补充 ASC-EVs 组中凋亡细胞显着减少。此外，通过蛋白质组学，我们检测到了每个治疗组的 ASC-EVs 和囊胚中的蛋白质。该分析显示，补充 ASC-EVs 组的蛋白质比例高于对照组（分别为 1,547 和 1,495）。基因分析证实，ASC-EVs 显示酪氨酸蛋白激酶 （SRC） 的高表达，而补充 ASC-EVs 的囊胚显示细胞周期蛋白依赖性激酶 1 （CDK1） 的表达较高。假设 SRC 激活蛋白激酶 B （AKT），从而抑制叉头盒 O 信号通路并激活 CDK1。 随后，CDK1 激活影响细胞周期，从而影响体外胚胎发育。ASC-EVs 促进线粒体活性，这对于囊胚的早期发育至关重要，对细胞凋亡的下调也至关重要。此外，ASC-EV 为猪囊胚提供 SRC，从而延长细胞周期。