Department of Urology, First Affiliated Hospital of Soochow University, Suzhou, China.
Department of Infectious Diseases, Children's Hospital of Soochow University, Suzhou, China.
Int J Artif Organs. 2022 Feb;45(2):181-192. doi: 10.1177/0391398820986809. Epub 2021 Jan 19.
Three-dimensional (3D) culture has been reported to increase the therapeutic potential of mesenchymal stem cells (MSCs). The present study assessed the therapeutic efficacy of extracellular vesicles (EVs) from 3D cultures of human placental MSCs (hPMSCs) for acute kidney injury (AKI).
The supernatants from monolayer culture (2D) and 3D culture of hPMSCs were ultra-centrifuged for EVs isolation. C57BL/6 male mice were submitted to 45 min bilateral ischemia of kidney, followed by renal intra-capsular administration of EVs within a 72 h reperfusion period. Histological, immunohistochemical, and ELISA analyses of kidney samples were performed to evaluate cell death and inflammation. Kidney function was evaluated by measuring serum creatinine and urea nitrogen. The miRNA expression profiles of EVs from 2D and 3D culture of hPMSCs were evaluated using miRNA microarray analysis.
The 3D culture of hPMSCs formed spheroids with different diameters depending on the cell density seeded. The hPMSCs produced significantly more EVs in 3D culture than in 2D culture. More importantly, injection of EVs from 3D culture of hPMSCs into mouse kidney with ischemia-reperfusion (I/R)-AKI was more beneficial in protecting from progression of I/R than those from 2D culture. The EVs from 3D culture of hPMSCs were more efficient against apoptosis and inflammation than those from 2D culture, which resulted in a reduction in tissue damage and amelioration of renal function. MicroRNA profiling analysis revealed that a set of microRNAs were significantly changed in EVs from 3D culture of hPMSCs, especially miR-93-5p.
The EVs from 3D culture of hPMSCs have therapeutic potential for I/R-AKI.
已有研究报道，三维（3D）培养可以提高间充质干细胞（MSCs）的治疗潜力。本研究评估了人胎盘间充质干细胞（hPMSCs）3D 培养的细胞外囊泡（EVs）治疗急性肾损伤（AKI）的疗效。
单层培养（2D）和 hPMSC 3D 培养的上清液经超速离心分离 EVs。C57BL/6 雄性小鼠进行双侧肾脏 45 分钟缺血，再灌注 72 小时内肾内囊内给予 EVs。对肾脏样本进行组织学、免疫组织化学和 ELISA 分析，以评估细胞死亡和炎症。通过测量血清肌酐和尿素氮评估肾功能。采用 miRNA 微阵列分析评估 hPMSC 2D 和 3D 培养的 EVs 的 miRNA 表达谱。
hPMSC 的 3D 培养形成了不同直径的球体，这取决于接种的细胞密度。与 2D 培养相比，hPMSC 在 3D 培养中产生的 EVs 显著增加。更重要的是，将来自 hPMSC 3D 培养的 EVs 注射到缺血再灌注（I/R）-AKI 小鼠的肾脏中，比注射来自 2D 培养的 EVs 更有利于保护免受 I/R 进展的影响。与 2D 培养相比，来自 hPMSC 3D 培养的 EVs 对细胞凋亡和炎症的抑制作用更强，这导致组织损伤减少和肾功能改善。miRNA 谱分析显示，hPMSC 3D 培养的 EVs 中一组 microRNAs 明显改变，特别是 miR-93-5p。
hPMSC 3D 培养的 EVs 具有治疗 I/R-AKI 的潜力。
