Tsubaki Toshiya, Chijimatsu Ryota, Takeda Taiga, Abe Maki, Ochiya Takahiro, Tsuji Shinsaku, Inoue Keita, Matsuzaki Tokio, Iwanaga Yasuhide, Omata Yasunori, Tanaka Sakae, Saito Taku
Orthopaedic Surgery, Sensory and Motor System Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-Ku, Tokyo, 113-8655 Japan.
Center for Comprehensive Genomic Medicine, Okayama University Hospital, 2-5-1, Shikada-Chou, Kita-Ku, Okayama, 700-8558 Japan.
Cytotechnology. 2025 Feb;77(1):15. doi: 10.1007/s10616-024-00675-6. Epub 2024 Dec 10.
Adipose-derived stem cells (ASCs) and their small extracellular vesicles (sEVs) hold significant potential for regenerative medicine due to their tissue repair capabilities. The microRNA (miRNA) content in sEVs varies depending on ASC status; however, the effects of aging and cell passage on miRNA profiles remain unclear. In this study, we examined the effects of donor age and cell expansion on ASC characteristics and transcriptome using ASCs obtained from three young and three old donors. Cell expansion significantly impaired stem cell properties, notably reducing proliferation and differentiation capacities. In contrast, donor age had minimal effects on ASCs. RNA sequencing (RNA-seq) revealed differences in gene expression related to stemness, phagocytosis, and metabolic processes influenced by cell expansion. To investigate miRNA variability, we performed small RNA-seq on sEVs collected from ASCs of all six donors. The miRNA profiles were influenced by donor age and cell passage. Interestingly, functional enrichment analysis indicated that advanced donor age and increased cell passage may enhance the production of miRNAs associated with organ development through various pathways. These findings suggest that donor age and cell expansion differentially influence ASC characteristics and sEV miRNA content, highlighting the need for disease-specific conditioning of ASCs to optimize the therapeutic effects of sEVs in clinical applications.
The online version contains supplementary material available at 10.1007/s10616-024-00675-6.
脂肪来源干细胞（ASCs）及其小细胞外囊泡（sEVs）因其组织修复能力在再生医学中具有巨大潜力。sEVs中的微小RNA（miRNA）含量因ASC状态而异；然而，衰老和传代对miRNA谱的影响仍不清楚。在本研究中，我们使用从三名年轻供体和三名老年供体获得的ASCs，研究了供体年龄和细胞传代对ASC特征和转录组的影响。细胞传代显著损害干细胞特性，特别是降低增殖和分化能力。相比之下，供体年龄对ASCs的影响最小。RNA测序（RNA-seq）揭示了与干性、吞噬作用和受细胞传代影响的代谢过程相关的基因表达差异。为了研究miRNA的变异性，我们对从所有六名供体的ASCs收集的sEVs进行了小RNA测序。miRNA谱受供体年龄和细胞传代的影响。有趣的是，功能富集分析表明，供体年龄增加和细胞传代增加可能通过各种途径增强与器官发育相关的miRNA的产生。这些发现表明，供体年龄和细胞传代对ASC特征和sEV miRNA含量有不同影响，突出了对ASCs进行疾病特异性处理以优化sEVs在临床应用中的治疗效果的必要性。
在线版本包含可在10.1007/s10616-024-00675-6获取的补充材料。
