Effects of cobalt chloride on the stem cell marker expression and osteogenic differentiation of stem cells from human exfoliated deciduous teeth
Effects of cobalt chloride on the stem cell marker expression and osteogenic differentiation of stem cells from human exfoliated deciduous teeth
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DOI：
10.1007/s12192-019-00981-5
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发表时间：
2019-05-01
影响因子：
3.8
通讯作者：
Zhao, Wei
中科院分区：
文献类型：
--
作者：
Chen, Yijing;Zhao, Qi;Zhao, Wei
来自人类脱落乳牙（SHED）的干细胞是组织工程和干细胞移植的一个有前途的来源。然而，长期的体外培养和扩增导致SHED的干性丧失，损害了它们的治疗益处。缺氧在控制间充质干细胞（MSCs）的干细胞行为中起着至关重要的作用。因此，本研究旨在研究氯化钴（CoCl 2），缺氧模拟剂，对干细胞标志物的表达和成骨分化的SHED的影响。SHED用或不用50或100 M CoCl 2培养。检测其增殖、凋亡、干细胞标志物表达、迁移能力和成骨分化。培养50和100 M的氯化钴增加缺氧诱导因子-1 α（HIF-1）的蛋白水平在SHED剂量依赖性的方式，而不诱导显着的细胞毒性。这种效应伴随着STRO-1（+）细胞比例的增加。CoCl 2以剂量依赖性方式显著增加干细胞标志物（OCT 4、NANOG、SOX 2和c-Myc）的表达。CoCl 2处理也促进了迁移能力。此外，SHEDs培养成骨培养基与氯化钴显示了剂量依赖性的碱性磷酸酶（ALP）活性和钙沉积的减少。成骨相关基因的表达也被CoCl 2抑制，尤其是在100-M CoCl 2组。结论：CoCl 2可增加SHEDs干细胞标志物的表达，抑制SHEDs的成骨分化。这些发现可能提供证据支持使用体外低氧环境模拟的氯化钴在辅助SHED的临床应用。
Stem cells from human exfoliated deciduous teeth (SHEDs) are a promising source for tissue engineering and stem cell transplantation. However, long-term in vitro culture and expansion lead to the loss of stemness of SHEDs, compromising their therapeutic benefits. Hypoxia plays an essential role in controlling the stem cell behavior of mesenchymal stem cells (MSCs). Thus, this study aimed to investigate the effects of cobalt chloride (CoCl2), a hypoxia-mimetic agent, on the stem cell marker expression and osteogenic differentiation of SHEDs. SHEDs were cultured with or without 50 or 100M CoCl2. Their proliferation, apoptosis, stem cell marker expression, migration ability, and osteogenic differentiation were examined. Culture with 50 and 100M CoCl2 increased the hypoxia-inducible factor-1 alpha (HIF-1) protein levels in a dose-dependent manner in SHEDs without inducing significant cytotoxicity. This effect was accompanied by an increase in the proportion of STRO-1(+) cells. CoCl2 significantly increased the expression of stem cell markers (OCT4, NANOG, SOX2, and c-Myc) in a dose-dependent manner. The migration ability was also promoted by CoCl2 treatment. Furthermore, SHEDs cultured in osteogenic medium with CoCl2 showed a dose-dependent reduction in alkaline phosphatase (ALP) activity and calcium deposition. The expression of osteogenic-related genes was also suppressed by CoCl2, especially in the 100-M CoCl2 group. In conclusion, CoCl2 increased the expression of stem cell markers and inhibited the osteogenic differentiation of SHEDs. These findings may provide evidence supporting the use of in vitro hypoxic environments mimicked by CoCl2 in assisting the clinical application of SHEDs.