The effects of short-term hypoxia on human mesenchymal stem cell proliferation, viability and p16(INK4A) mRNA expression: Investigation using a simple hypoxic culture system with a deoxidizing agent.
The effects of short-term hypoxia on human mesenchymal stem cell proliferation, viability and p16(INK4A) mRNA expression: Investigation using a simple hypoxic culture system with a deoxidizing agent.
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DOI：
10.46582/jsrm.1101005
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发表时间：
2015
影响因子：
2.7
通讯作者：
Kuroki H
中科院分区：
文献类型：
--
作者：
Ito A;Aoyama T;Yoshizawa M;Nagai M;Tajino J;Yamaguchi S;Iijima H;Zhang X;Kuroki H
低氧环境被认为对于干细胞中的干细胞性的维持和抑制细胞衰老是重要的。因此，在细胞扩增和/或诱导预期分化期间诱导缺氧条件。然而，这些条件的诱导需要专门装备的低氧室和昂贵的气体混合物，这是昂贵的和占用空间的。由于这些限制，在细胞运输过程中无法提供适当的缺氧条件，而再生医学越来越需要这种条件。因此，需要一种简单且经济的培养系统。本研究的目的是调查短期缺氧条件对人骨髓间充质干细胞（MSC）的增殖，活力和衰老的影响，利用培养系统（CulturePal-Zero和CulturePal-Five），一种新型的和简单的缺氧培养系统，内置的脱氧剂。观察到CulturePal-Zero中的O2浓度在1小时内降低至<0.1%，在CulturePal-Five系统中在24小时内降低至5%。在这些低氧条件下的细胞增殖显示在5%O2浓度下急剧增加，并且即使在严重低氧条件下（<0.1%O2）长达72 h也没有观察到明显的细胞死亡。p16 INK 4A（细胞衰老标志物）mRNA表达在低氧条件下保持至72 h，但在常氧条件下上调。有趣的是，p16 INK 4A的表达与O2浓度成比例地改变。这些结果表明，短期低氧条件下，在约5%的O2浓度，将是适合于促进细胞增殖和抑制细胞衰老，而不会加重MSC的活力。因此，培养系统可能适合于在干细胞研究和运输中提供适当的低氧条件。
A hypoxic environment is thought to be important for the maintenance of stemness and suppressing cell senescence, in stem cells. Therefore, a hypoxic condition is induced during cell expansion and/or induction of intended differentiation. However, the induction of these conditions requires a specially equipped hypoxia chamber and expensive gas mixtures, which are expensive and space-consuming. Owing to these restrictions, appropriate hypoxic conditions cannot be provided during cell transportation, which is increasingly required for regenerative medicine. Hence, a simple and economical culture system is required. The purpose of this study was to investigate the effects of short-term hypoxic conditions on human mesenchymal stem cell (MSC) proliferation, viability, and senescence, utilizing the CulturePal system (CulturePal-Zero and CulturePal-Five), a novel and simple hypoxic culture system with a built-in deoxidizing agent. The O2 concentration in the CulturePal-Zero was observed to reduce to <0.1% within 1 h, and to 5% within 24h in the CulturePal-Five system. Cell proliferation under these hypoxic conditions showed a sharp increase at 5% O2 concentration, and no noticeable cell death was observed even at severe hypoxic conditions (<0.1% O2) up to 72h. The p16INK4A (cell senescence marker) mRNA expression was retained under hypoxic conditions up to 72h, but it was up-regulated under normoxic conditions. Interestingly, the p16INK4A expression altered proportionately to the O2 concentration. These results indicated that the short-term hypoxic condition, at an approximate O2 concentration of 5%, would be suitable for promoting cell proliferation and repressing cell senescence, without aggravating the MSC viability. Therefore, the CulturePal systems may be suitable for providing an appropriate hypoxic condition in stem cell research and transportation.