脂肪干细胞细胞外囊泡所含外源蛋白的胚胎营养效应

IF 6.3 Q1 AGRICULTURE, DAIRY & ANIMAL SCIENCE Journal of Animal Science and Biotechnology Pub Date : 2024-11-03 DOI:10.1186/s40104-024-01106-4
Seonggyu Bang, Ahmad Yar Qamar, Sung Ho Yun, Na-Yeon Gu, Heyyoung Kim, Ayeong Han, Heejae Kang, Hye Sun Park, Seung Ii Kim, Islam M Saadeldin, Sanghoon Lee, Jongki Cho
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引用次数: 0

摘要

背景:细胞外囊泡(EVs)通过向周围细胞输送特定的蛋白质和核酸来调节细胞代谢和各种生物过程。我们的目的是研究源自脂肪源性干细胞(ASCs)的EVs所含货物对猪胚胎发育的影响:方法:从猪脂肪组织中分离出 ASCs,并通过流式细胞术使用 ASC 特异性标记物对其进行鉴定。随后从已建立的 ASCs 的条件培养基中提取 EVs。将这些EVs加入猪胚胎的体外培养环境中,观察胚胎发育的质量改善情况。此外,还对 EVs 中的蛋白质进行了分析,以研究其潜在机制:结果:我们观察到,与对照组相比,添加 ASC-EVs 的早期胚胎囊胚发育率更高,线粒体活性也更强(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 缺口标记(TUNEL)检测也显示,ASC-EVs 补充组的凋亡细胞显著减少。此外,我们还通过蛋白质组学检测了各处理组 ASC-EV 和囊胚中的蛋白质。这项分析表明,补充 ASC-EVs 的组比对照组的蛋白质比例更高(分别为 1547 对 1495)。基因分析证实,ASC-EVs 显示出酪氨酸蛋白激酶(SRC)的高表达,而补充 ASC-EVs 的囊胚显示出细胞周期蛋白依赖性激酶 1(CDK1)的高表达。据推测,SRC 可激活蛋白激酶 B(AKT),而 AKT 可抑制叉头盒 O 信号通路并激活 CDK1。随后,CDK1的激活会影响细胞周期,从而影响体外胚胎发育:结论:ASC-EV 可促进线粒体活性,这对囊胚的早期发育至关重要,也是下调细胞凋亡的关键。此外,ASC-EV 还能为猪囊胚提供 SRC,从而延长细胞周期。
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Embryotrophic effect of exogenous protein contained adipose-derived stem cell extracellular vesicles.

Background: 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.

Methods: 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.

Results: 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.

Conclusion: 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.

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来源期刊
CiteScore
10.30
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发文量
822
期刊最新文献
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