Graduate School of International Agricultural Technology, Seoul National University, South Korea.
Nissan Chemical Corporation, Japan.
Biomaterials. 2021 Apr;271:120742. doi: 10.1016/j.biomaterials.2021.120742. Epub 2021 Mar 1.
Mesenchymal stem cell (MSC) transplantation is a promising therapy for regenerative medicine. However, MSCs grown under two-dimensional (2D) culture conditions differ significantly in cell shape from those in the body, with downregulated stemness genes and secretion of paracrine factors. Here, we evaluated the effect of 3D culture using Cellhesion VP, a water-insoluble material composed of chitin-based polysaccharide fibers, on the characteristics of human Wharton's jelly-derived MSCs (hMSCs). Cellhesion VP significantly increased cell proliferation after retrieval. Transcriptome analyses suggested that genes involved in cell stemness, migration ability, and extracellular vesicle (EV) production were enhanced by 3D culture. Subsequent biochemical analyses showed that the expression levels of stemness genes including OCT4, NANOG, and SSEA4 were upregulated and migration capacity was elevated in 3D-cultured hMSCs. In addition, EV production was significantly elevated in 3D cells, which contained a distinct protein profile from 2D cells. Gene and drug connectivity analyses revealed that the 2D and 3D EVs had similar functions as immunomodulators; however, 3D EVs had completely distinct therapeutic profiles for various infectious and metabolic diseases based on activation of disease-associated signaling pathways. Therefore, EVs from Cellhesion VP-primed hMSCs offer a new treatment for immune and metabolic diseases.
间充质干细胞 (MSC) 移植是再生医学有前途的治疗方法。然而，在二维 (2D) 培养条件下生长的 MSC 在细胞形状上与体内的 MSC 有很大的不同，其干性基因下调，旁分泌因子的分泌减少。在这里，我们评估了使用 Cellhesion VP 进行 3D 培养对人脐带来源 MSC (hMSC) 特性的影响。Cellhesion VP 在回收后显著增加细胞增殖。转录组分析表明，3D 培养增强了与细胞干性、迁移能力和细胞外囊泡 (EV) 产生相关的基因。随后的生化分析表明，3D 培养的 hMSC 中干性基因（包括 OCT4、NANOG 和 SSEA4）的表达上调，迁移能力增强。此外，3D 细胞中 EV 的产生显著增加，与 2D 细胞中的 EV 相比，其蛋白谱明显不同。基因和药物连接性分析表明，2D 和 3D EV 具有相似的免疫调节功能；然而，3D EV 根据激活与疾病相关的信号通路，对各种感染性和代谢性疾病具有完全不同的治疗谱。因此，来自 Cellhesion VP 预激活 hMSC 的 EV 为免疫和代谢性疾病提供了一种新的治疗方法。
