Key Laboratory of Interdisciplinary Research, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
University of Chinese Academy of Sciences, Beijing, 100049, China.
Stem Cell Res Ther. 2022 Sep 5;13(1):449. doi: 10.1186/s13287-022-03142-1.
Increasing studies have reported the therapeutic effect of mesenchymal stem cell (MSC)-derived exosomes by which protein and miRNA are clearly characterized. However, the proteomics and miRNA profiles of exosomes derived from human embryonic stem cells (hESCs) and human-induced pluripotent stem cells (hiPSCs) remain unclear.
In this study, we isolated exosomes from hESCs, hiPSCs, and human umbilical cord mesenchymal stem cells (hUC-MSCs) via classic ultracentrifugation and a 0.22-μm filter, followed by the conservative identification. Tandem mass tag labeling and label-free relative peptide quantification together defined their proteomics. High-throughput sequencing was performed to determine miRNA profiles. Then, we conducted a bioinformatics analysis to identify the dominant biological processes and pathways modulated by exosome cargos. Finally, the western blot and RT-qPCR were performed to detect the actual loads of proteins and miRNAs in three types of exosomes.
Based on our study, the cargos from three types of exosomes contribute to sophisticated biological processes. In comparison, hESC exosomes (hESC-Exos) were superior in regulating development, metabolism, and anti-aging, and hiPSC exosomes (hiPSC-Exos) had similar biological functions as hESC-Exos, whereas hUC-MSCs exosomes (hUC-MSC-Exos) contributed more to immune regulation.
The data presented in our study help define the protein and miRNA landscapes of three exosomes, predict their biological functions via systematic and comprehensive network analysis at the system level, and reveal their respective potential applications in different fields so as to optimize exosome selection in preclinical and clinical trials.
越来越多的研究报告了间充质干细胞（MSC）衍生的外泌体的治疗效果，其中蛋白质和 miRNA 得到了明确的表征。然而，人胚胎干细胞（hESC）和人诱导多能干细胞（hiPSC）衍生的外泌体的蛋白质组学和 miRNA 图谱仍不清楚。
在这项研究中，我们通过经典的超速离心和 0.22μm 滤器从 hESC、hiPSC 和人脐带间充质干细胞（hUC-MSC）中分离出外泌体，随后进行保守鉴定。串联质量标签标记和无标记相对肽定量一起定义了它们的蛋白质组学。高通量测序用于确定 miRNA 图谱。然后，我们进行了生物信息学分析，以确定外泌体货物调节的主要生物学过程和途径。最后，western blot 和 RT-qPCR 用于检测三种类型外泌体中的蛋白质和 miRNA 的实际负载。
基于我们的研究，三种类型的外泌体的货物有助于复杂的生物学过程。相比之下，hESC 外泌体（hESC-Exos）在调节发育、代谢和抗衰老方面表现更为出色，hiPSC 外泌体（hiPSC-Exos）与 hESC-Exos 具有相似的生物学功能，而 hUC-MSC 外泌体（hUC-MSC-Exos）则更有助于免疫调节。
本研究中提供的数据有助于定义三种外泌体的蛋白质和 miRNA 图谱，通过系统和全面的网络分析在系统水平上预测它们的生物学功能，并揭示它们各自在不同领域的潜在应用，从而优化临床前和临床试验中外泌体的选择。
