Alwohoush Enas, Ismail Mohammad A, Al-Kurdi Ban, Barham Raghda, Al Hadidi Sabal, Awidi Abdalla, Ababneh Nidaa A
Cell Therapy Center, the University of Jordan, Amman, Jordan.
Hemostasis and Thrombosis Laboratory, School of Medicine, the University of Jordan, Amman, Jordan.
Heliyon. 2024 Oct 2;10(19):e38857. doi: 10.1016/j.heliyon.2024.e38857. eCollection 2024 Oct 15.
Although mesenchymal stem cells (MSCs) are extensively applied in the regenerative field, the majority of MSCs die after a few weeks of transplantation. Therefore, hypoxia pre-conditioning is a crucial step in increasing the MSCs' tolerance to physiological conditions. Meanwhile, induced pluripotent stem cell-derived MSCs (iMSCs) were proposed as a possible alternative to MSCs, and recently, the interest is growing in applying iMSCs in the regenerative field. This study examined the effect of hypoxia pre-conditioning on the proliferation, viability, and differentiation of iMSCs. Both iMSCs and MSCs were subjected to two rounds of severe short-term hypoxia (1 % O for 24h). After that, iMSCs and MSCs were characterized by testing their surface markers' expression, proliferation, viability, oxidative stress, and differentiation potential. Our findings revealed that hypoxia did not have a consistent effect among all the analyzed lines: the severe short-term hypoxia (1 % O) reduced iMSCs proliferation, cell viability, and MMP while showing a benign effect on surface markers expression, colony formation, ROS accumulation, and osteogenic and adipogenic differentiation. Though hypoxia adversely affected iMSCs' proliferation, this does not necessarily mean that hypoxia is harmful to iMSCs; on the contrary, our results suggest that short-term hypoxia might have a beneficial long-term effect on the proliferation of iMSCs. Thus, the effect of hypoxia on proliferation, viability, and differentiation should also be tested after a long recovery period from iMSCs. Our next step will be to test the effect of hypoxia for a longer period besides uncovering the changes in the expression profile of hypoxic iMSCs.
尽管间充质干细胞（MSCs）在再生领域得到了广泛应用，但大多数MSCs在移植几周后就会死亡。因此，缺氧预处理是提高MSCs对生理条件耐受性的关键步骤。同时，诱导多能干细胞衍生的间充质干细胞（iMSCs）被提议作为MSCs的一种可能替代物，最近，将iMSCs应用于再生领域的兴趣日益浓厚。本研究考察了缺氧预处理对iMSCs增殖、活力和分化的影响。iMSCs和MSCs均经历两轮严重短期缺氧（1%氧气，持续24小时）。之后，通过检测iMSCs和MSCs的表面标志物表达、增殖、活力、氧化应激和分化潜能对其进行表征。我们的研究结果显示，缺氧在所有分析的细胞系中并没有一致的影响：严重短期缺氧（1%氧气）降低了iMSCs的增殖、细胞活力和线粒体膜电位，同时对表面标志物表达、集落形成、活性氧积累以及成骨和成脂分化显示出良性影响。尽管缺氧对iMSCs的增殖有不利影响，但这并不一定意味着缺氧对iMSCs有害；相反，我们的结果表明短期缺氧可能对iMSCs的增殖有长期有益影响。因此，在iMSCs经过长时间恢复后，也应测试缺氧对其增殖、活力和分化的影响。我们的下一步将是除了揭示缺氧iMSCs表达谱的变化外，测试更长时间缺氧的影响。
