Haematologica. 2013 Oct;98(10):1609-16. doi: 10.3324/haematol.2013.092700. Epub 2013 Aug 23.
Previous reports demonstrated a relationship between proliferation potential and trilineage differentiation in mesenchymal stromal cell-derived clones generated using plastic adherence (PA-MSCs). However, there are no reports presenting a clonal analysis of the proliferative potential, differentiation potential and allosuppressive effects of human mesenchymal stromal cell subsets. In this study, we performed a clonal analysis of mesenchymal stromal cells generated from human CD271(+) bone marrow mononuclear cells (CD271-MSCs). After transfection with the gene encoding green fluorescent protein, the cells were single-cell sorted and cultured for 2-4 weeks. A population doubling analysis demonstrated that 25% of CD271-MSC clones are fast-proliferating clones compared to only 10% of PA-MSC clones. Evaluation of the allosuppressive potential demonstrated that 81.8% of CD271-MSC clones were highly allosuppressive compared to only 58% of PA-MSC clones. However, no consistent correlation was observed between allosuppression and proliferative potential. Prostaglandin E2 levels were positively correlated with the allosuppressive activity of individual clones, suggesting that this molecule may be a useful predictive biomarker for the allosuppressive potential of mesenchymal stromal cells. In contrast, inhibitory studies of indoleamine 2,3 dioxygenase indicated that none of the clones used this enzyme to mediate their allosuppressive effect. Differentiation studies revealed the presence of tripotent, bipotent and unipotent CD271-MSC and PA-MSC clones which suppressed the allogeneic reaction to differing extents in vitro. In conclusion, our findings demonstrate differences between CD271-MSCs and PA-MSCs and indicate that neither proliferation potential nor differentiation potential represents a consistent predictive parameter for the immunomodulatory effects of either type of mesenchymal stromal cells.
先前的报告表明，在使用塑料贴附（PA-MSCs）生成的间充质基质细胞衍生克隆中，增殖潜能与三系分化之间存在关系。然而，目前尚无关于人类间充质基质细胞亚群的增殖潜能、分化潜能和免疫抑制作用的克隆分析的报道。在这项研究中，我们对来源于人 CD271+骨髓单核细胞（CD271-MSCs）的间充质基质细胞进行了克隆分析。转染编码绿色荧光蛋白的基因后，对细胞进行单细胞分选并培养 2-4 周。群体倍增分析表明，与仅 10%的 PA-MSC 克隆相比，25%的 CD271-MSC 克隆为快速增殖克隆。对免疫抑制潜能的评估表明，81.8%的 CD271-MSC 克隆具有高度免疫抑制作用，而仅 58%的 PA-MSC 克隆具有免疫抑制作用。然而，在免疫抑制作用和增殖潜能之间未观察到一致的相关性。前列腺素 E2 水平与单个克隆的免疫抑制活性呈正相关，表明该分子可能是间充质基质细胞免疫抑制潜能的有用预测生物标志物。相反，吲哚胺 2,3 双加氧酶的抑制研究表明，使用的克隆均未利用该酶介导其免疫抑制作用。分化研究揭示了存在三潜能、双潜能和单潜能的 CD271-MSC 和 PA-MSC 克隆，它们在体外以不同程度抑制同种异体反应。总之，我们的研究结果表明 CD271-MSCs 和 PA-MSCs 之间存在差异，并且增殖潜能和分化潜能均不能代表两种类型的间充质基质细胞的免疫调节作用的一致预测参数。
