Center for Craniofacial Molecular Biology, The Herman Ostrow School of Dentistry of University of Southern California, Los Angeles, California 90033, USA.
Stem Cells Dev. 2012 Apr 10;21(6):937-47. doi: 10.1089/scd.2011.0252. Epub 2011 Jul 28.
Mesenchymal stem cells (MSCs) are capable of regenerative and immunomodulatory functions in cell-based therapies in a variety of human diseases and injuries; however, their therapeutic efficacy and potential side effects remain major obstacles in clinical applications. We report here a 3D spheroid culture approach to optimize stem cell properties and therapeutic effects of human gingiva-derived mesenchymal stem cells (GMSCs) in mitigation of experimental oral mucositis. Under growth condition of ultra-low attachment, GMSCs spontaneously aggregated into 3D spheroids and exhibited distinct early stem cell phenotype characterized by elevated expression Stro-1 and CXC chemokine receptor 4 (CXCR-4) as well as OCT-4 and Nanog, 2 important transcriptional factors relevant to stem cell properties, and decreased expression of MSC-associated markers, including CD29, CD90, and CD105. Functionally, spheroid GMSCs are capable of enhanced multipotency and augmented secretion of several chemokines and cytokines relevant to cell migration, survival, and angiogenesis. More importantly, spheroid GMSCs expressed increased levels of reactive oxygen species, hypoxia-inducible factor (HIF)-1 and -2α, and manganese superoxide dismutase, which correlated with improved resistance to oxidative stress-induced apoptosis. Using an in vivo murine model of chemotherapy-induced oral mucositis, we demonstrated that spheroid-derived GMSCs possessed better therapeutic efficacy than their adherent cells in reversing body weight loss and promoting the regeneration of disrupted epithelial lining of the mucositic tongues. These findings suggest that 3D spheroid culture allows early stemness preservation and potentially precondition GMSCs for enhanced mitigation of oral mucositis.
间充质干细胞（MSCs）在多种人类疾病和损伤的细胞治疗中具有再生和免疫调节功能；然而，其治疗效果和潜在的副作用仍然是临床应用的主要障碍。我们在此报告一种 3D 球体培养方法，以优化人牙龈来源间充质干细胞（GMSCs）的干细胞特性和治疗效果，减轻实验性口腔黏膜炎。在超低附着生长条件下，GMSCs自发聚集形成 3D 球体，并表现出明显的早期干细胞表型，特征为 Stro-1 和 CXC 趋化因子受体 4（CXCR-4）以及 OCT-4 和 Nanog 的表达升高，这 2 个转录因子与干细胞特性相关，以及 MSC 相关标志物，包括 CD29、CD90 和 CD105 的表达降低。功能上，球体 GMSCs 具有增强的多能性和几种与细胞迁移、存活和血管生成相关的趋化因子和细胞因子的分泌增强。更重要的是，球体 GMSCs 表达增加的活性氧、缺氧诱导因子（HIF）-1 和 -2α 以及锰超氧化物歧化酶，与提高对氧化应激诱导的细胞凋亡的抗性相关。使用化疗诱导的口腔黏膜炎的体内小鼠模型，我们证明了球体衍生的 GMSCs 比其贴壁细胞具有更好的治疗效果，可逆转体重减轻并促进黏膜炎舌上皮层的破坏再生。这些发现表明，3D 球体培养允许早期干细胞特性的保存，并可能为减轻口腔黏膜炎提供 GMSCs 的增强预处理。
