Aggregation of human mesenchymal stromal cells (MSCs) into 3D spheroids enhances their antiinflammatory properties
Aggregation of human mesenchymal stromal cells (MSCs) into 3D spheroids enhances their antiinflammatory properties
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DOI：
10.1073/pnas.1008117107
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发表时间：
2010-08-03
影响因子：
11.1
通讯作者：
Prockop, Darwin J.
中科院分区：
文献类型：
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作者：
Bartosh, Thomas J.;Ylostalo, Joni H.;Prockop, Darwin J.
以前的报道表明，作为3D聚集体或作为球状体培养可以增加称为间充质干细胞或多能间充质基质细胞（MSC）的成体干/祖细胞的治疗潜力。在这里，我们使用悬滴方案将人MSC（hMSC）制备为最大表达TNF α刺激的基因/蛋白6（TSG-6）的球体，TNF α刺激的基因/蛋白6（TSG-6）是在静脉输注后被困在肺中的hMSC以高水平表达的TNF α蛋白，并且在很大程度上解释了hMSC在患有心肌梗死的小鼠中的有益作用。发现球状体hMSCs的性质关键取决于培养条件。在表达TSG-6的最佳条件下，hMSCs还表达高水平的斯钙素-1，一种具有抗凋亡和抗凋亡特性的蛋白质。此外，它们表达高水平的三种抗癌蛋白：IL-24、TNF α相关凋亡诱导配体和CD 82。在与LPS激活的巨噬细胞共培养系统和腹膜炎小鼠模型中，球状体hMSCs比贴壁单层培养的hMSCs更有效地抑制炎症反应。此外，球状体hMSC的体积约为贴壁培养的hMSC体积的四分之一。显然，结果是，静脉输注后大量细胞通过肺运输，并在脾、肝、肾和心脏中回收。这些数据表明，球状体hMSC可能比贴壁培养的hMSC更有效地治疗以无菌组织损伤和未解决的炎症为特征的疾病以及对化疗剂敏感的一些癌症。
Previous reports suggested that culture as 3D aggregates or as spheroids can increase the therapeutic potential of the adult stem/progenitor cells referred to as mesenchymal stem cells or multipotent mesenchymal stromal cells (MSCs). Here we used a hanging drop protocol to prepare human MSCs (hMSCs) as spheroids that maximally expressed TNF alpha stimulated gene/protein 6 (TSG-6), the antiinflammatory protein that was expressed at high levels by hMSCs trapped in the lung after i.v. infusion and that largely explained the beneficial effects of hMSCs in mice with myocardial infarcts. The properties of spheroid hMSCs were found to depend critically on the culture conditions. Under optimal conditions for expression of TSG-6, the hMSCs also expressed high levels of stanniocalcin-1, a protein with both antiinflammatory and antiapoptotic properties. In addition, they expressed high levels of three anticancer proteins: IL-24, TNF alpha-related apoptosis inducing ligand, and CD82. The spheroid hMSCs were more effective than hMSCs from adherent monolayer cultures in suppressing inflammatory responses in a coculture system with LPS-activated macrophages and in a mouse model for peritonitis. In addition, the spheroid hMSCs were about one-fourth the volume of hMSCs from adherent cultures. Apparently as a result, larger numbers of the cells trafficked through the lung after i.v. infusion and were recovered in spleen, liver, kidney, and heart. The data suggest that spheroid hMSCs may be more effective than hMSCs from adherent cultures in therapies for diseases characterized by sterile tissue injury and unresolved inflammation and for some cancers that are sensitive to antiinflammatory agents.