Choi Wankyu, Kwon Soon-Jae, Jin Hye Jin, Jeong Sang Young, Choi Soo Jin, Oh Wonil, Yang Yoon Sun, Jeon Hong Bae, Jeon Eun Su
Biomedical Research Institute, R&D Center, MEDIPOST Co., Ltd., 21 Daewangpangyo-ro 644beon-gil, Bundang-gu, Seongnam-Si, Gyeonggi-do, 13494, Republic of Korea.
Clin Transl Med. 2017 Oct 10;6(1):38. doi: 10.1186/s40169-017-0168-z.
Mesenchymal stem cells (MSCs) have broad-spectrum therapeutic effects in various diseases, and thus have many clinical applications. However, it is difficult to produce sufficient numbers of MSCs for clinical use, and improved culture systems are required. Here, we report the effects of calcium (Ca) and hypoxia on the proliferation of human umbilical cord blood-derived MSCs (hUCB-MSCs). In addition, we determined the optimal conditions of these two factors for the large-scale culture of hUCB-MSCs.
hUCB-MSCs were maintained under hypoxic conditions (3% O) with 1.8 mM Ca during long-term culture, and their proliferation was evaluated. To characterize the underlying mechanisms, the effects on hypoxia-inducible factor (HIF)-1α and the extracellular signal-regulated kinase (ERK) signaling pathways were investigated. The therapeutic effects in a mouse emphysema model were analyzed and compared with those of naive MSCs.
The proliferation of Ca/hypoxia-treated hUCB-MSCs was increased compared with that observed using either calcium or hypoxia culture alone, without loss of stem cell marker expression or differentiation ability. The enhancement of the proliferation capacity of hUCB-MSCs by the synergistic effects of Ca and hypoxia was dependent on the expression of HIF-1α and the ERK signaling pathway. The proliferation of Ca/hypoxia-treated hUCB-MSCs resulted in a delayed senescence phenotype and increased the expression levels of stemness genes such as Oct4 and Nanog compared to those observed in conventional culture conditions. In addition, Ca/hypoxia-treated MSCs transplantation in the mouse emphysema model showed the same therapeutic effects as observed with naive MSCs.
These findings suggest that a Ca/hypoxia-based expansion system has applications for the large-scale production of MSCs for therapeutic purposes.
间充质干细胞（MSCs）在多种疾病中具有广谱治疗作用，因此有许多临床应用。然而，难以生产出足够数量的MSCs用于临床，需要改进培养系统。在此，我们报告钙（Ca）和缺氧对人脐带血来源的间充质干细胞（hUCB-MSCs）增殖的影响。此外，我们确定了这两个因素用于hUCB-MSCs大规模培养的最佳条件。
在长期培养过程中，将hUCB-MSCs置于含1.8 mM Ca的低氧条件（3% O）下，并评估其增殖情况。为了阐明潜在机制，研究了对缺氧诱导因子（HIF）-1α和细胞外信号调节激酶（ERK）信号通路的影响。分析了在小鼠肺气肿模型中的治疗效果，并与未处理的MSCs进行比较。
与单独使用钙或缺氧培养相比，经Ca/缺氧处理的hUCB-MSCs的增殖增加，且未丧失干细胞标志物表达或分化能力。Ca和缺氧的协同作用增强hUCB-MSCs增殖能力依赖于HIF-1α的表达和ERK信号通路。与传统培养条件相比，经Ca/缺氧处理的hUCB-MSCs的增殖导致衰老表型延迟，并增加了Oct4和Nanog等干性基因的表达水平。此外，在小鼠肺气肿模型中移植经Ca/缺氧处理的MSCs显示出与未处理的MSCs相同的治疗效果。
这些发现表明，基于Ca/缺氧的扩增系统可用于大规模生产用于治疗目的的MSCs。
