United States Army Institute of Surgical Research, San Antonio, TX 78234, USA.
Oak Ridge Institute for Science and Education, Oak Ridge, TN 37830, USA.
Cells. 2019 May 15;8(5):462. doi: 10.3390/cells8050462.
Multipotent mesenchymal stromal cells (MSCs) have emerged as potent therapeutic agents for multiple indications. However, recent evidence indicates that MSC function is compromised in the physiological post-injury milieu. In this study, bone marrow (BM)- and adipose-derived (AD)-MSCs were preconditioned in hypoxia with or without inflammatory mediators to potentiate their immunotherapeutic function in preparation for in vivo delivery. Human MSCs were cultured for 48 hours in either normoxia (21% O) or hypoxia (2% O) with or without the addition of Cytomix, thus creating 4 groups: 1) normoxia (21%); 2) Cytomix-normoxia (+21%); 3) hypoxia (2%); and 4) Cytomix-hypoxia (+2%). The 4 MSC groups were subjected to comprehensive evaluation of their characteristics and function. Preconditioning did not alter common MSC surface markers; nonetheless, Cytomix treatment triggered an increase in tissue factor (TF) expression. Moreover, the BM-MSCs and AD-MSCs from the +2% group were not able to differentiate to chondrocytes and osteoblasts, respectively. Following Cytomix preconditioning, the metabolism of MSCs was significantly increased while viability was decreased in AD-MSCs, but not in BM-MSCs. MSCs from both tissues showed a significant upregulation of key anti-inflammatory genes, increased secretion of IL-1 receptor antagonist (RA), and enhanced suppression of T-cell proliferation following the Cytomix treatment. Similarly, following a lipopolysaccharide challenge, the Cytomix-treated MSCs suppressed TNF-α secretion, while promoting the production of IL-10 and IL-1RA. These preconditioning approaches facilitate the production of MSCs with robust anti-inflammatory properties. AD-MSCs preconditioned with Cytomix under normoxia appear to be the most promising therapeutic candidates; however, safety concerns, such as thrombogenic disposition of cells due to TF expression, should be carefully considered prior to clinical translation.
多能间充质基质细胞 (MSCs) 已成为多种适应证的有效治疗药物。然而，最近的证据表明，MSC 的功能在生理损伤环境中受到损害。在这项研究中，骨髓 (BM) 和脂肪组织 (AD) 来源的 MSC 分别在缺氧条件下或缺氧条件下联合炎症介质进行预处理，以增强其在体内递送前的免疫治疗功能。将人 MSC 在常氧 (21% O) 或缺氧 (2% O) 下培养 48 小时，或在添加 Cytomix 的情况下培养，从而创建了 4 个组：1) 常氧 (21%)；2) Cytomix-常氧 (+21%)；3) 缺氧 (2%)；和 4) Cytomix-缺氧 (+2%)。将这 4 组 MSC 进行了全面评估，以确定其特性和功能。预处理并没有改变常见的 MSC 表面标志物；然而，Cytomix 处理会触发组织因子 (TF) 表达的增加。此外，来自 +2% 组的 BM-MSCs 和 AD-MSCs 分别不能分化为软骨细胞和成骨细胞。经过 Cytomix 预处理后，AD-MSCs 的代谢明显增加，而 AD-MSCs 的活力下降，但 BM-MSCs 则没有。两种组织来源的 MSC 都显著上调了关键抗炎基因的表达，增加了白细胞介素-1 受体拮抗剂 (RA) 的分泌，并增强了对 T 细胞增殖的抑制作用。同样，在脂多糖刺激后，经 Cytomix 处理的 MSC 抑制 TNF-α 的分泌，同时促进 IL-10 和 IL-1RA 的产生。这些预处理方法有助于产生具有强大抗炎特性的 MSC。在常氧条件下用 Cytomix 预处理的 AD-MSCs 似乎是最有前途的治疗候选者；然而，在进行临床转化之前，应该仔细考虑细胞 TF 表达导致的血栓形成倾向等安全性问题。
