摘要
目的:探讨不同性别来源人胚胎干细胞外泌体(human embryonic stem cell-derived exosome,hESC-exo)和人羊膜间充质干细胞外泌体(human amniotic mesenchymal stem cell-derived exosome,hAMSC-exo)在蛋白质组学和转录组学层面的差异。方法:构建符合外泌体研究的培养体系,通过超速离心分别从男女株人胚胎干细胞(human embryonic stem cell,hESC)和人羊膜间充质干细胞(human amniotic mesenchymal stem cell,h AMSC)中获得质量符合研究要求的外泌体(exosome,exo),采用液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)分析来定义蛋白质组学表征,进行高通量测序以确定微小RNA(microRNA,miRNA)谱。通过生物信息学分析确定外泌体调控的主要生物学过程和通路。结果:在蛋白质组学和转录组学层面,hAMSC-exo男株与女株表征类似,而h ESC-exo男株与女株间存在较大差异。miRNA图谱和蛋白质组分析揭示了hESC-exo主要与发育、代谢相关,hAMSC-exo主要与免疫、代谢相关。结论:通过蛋白质组学和miRNA测序对不同性别来源的hAMSC-exo和hESC-exo进行了系统分析,预测它们的生物学功能,揭示其在不同领域的应用潜力,为临床前和临床试验中选择更优来源细胞提供新的视角。
Objective:To investigate proteomic and transcriptomic differences between exosomes derived from sex-specific human embryonic stem cells(hESC-exo)and human amniotic mesenchymal stem cells(hAMSC-exo)at the molecular level.Methods:A defined cell culture system optimized for exosome research was established.High-quality exosomes were isolated via ultracentrifugation from male and female lines of hESCs and hAMSCs.Proteomic characterization was performed using liquid chromatography-tandem mass spectrometry(LC-MS/MS),while microRNA(miRNA)profiles were determined by high-throughput sequencing.Bioinformatics analysis subsequently identified key biological processes and pathways regulated by the exosomes.Results:At both proteomic and transcriptomic levels,exosomes derived from male and female hAMSCs exhibited similar molecular profiles,whereas those originating from male and female hESCs showed significant sex-based differences.Integrated miRNA profiling and proteomic analysis revealed that hESC-exo were primarily associated with developmental and metabolic processes,while hAMSC-exo predominantly participated in immune and metabolic functions.Conclusion:Through systematic proteomic and miRNA sequencing analyses of sex-specific exosomes derived from hAMSCs and hESCs,this study predicted their biological functions and revealed their potential applications across multiple domains.These findings provide novel perspectives for selecting optimal cell sources in preclinical and clinical trials.
作者
黄蓓蓓
宁松
吴慧敏
覃莲菊
刁飞扬
HUANG Beibei;NING Song;WU Huimin;QIN Lianju;DIAO Feiyang(State Key Laboratory of Reproductive Medicine and Offspring Health,Nanjing Medical University,Nanjing 211103;Clinical Center of Reproductive Medicine,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210036,China)
出处
《南京医科大学学报(自然科学版)》
北大核心
2026年第2期188-201,共14页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家重点研发计划(2022YFC2702505)。
