Effects of osmotic and cold shock on adherent human mesenchymal stem cells during cryopreservation.
Effects of osmotic and cold shock on adherent human mesenchymal stem cells during cryopreservation.
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DOI：
10.1016/j.jbiotec.2012.09.004
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发表时间：
2012-12
影响因子：
4.1
通讯作者：
Xia Xu;Yang Liu;Z. Cui;Yuping Wei;Liang Zhang
中科院分区：
文献类型：
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作者：
Xia Xu;Yang Liu;Z. Cui;Yuping Wei;Liang Zhang
冷冻保存是长期储存细胞基质系统的最实用的方法之一，以确保组织工程，干细胞治疗和药物测试的现成可用性。本研究的目的是探讨渗透压和冷休克引起的冷冻保护剂的添加/删除和冷冻过程中对细胞活力，细胞内的特性，如丝状肌动蛋白分布，线粒体定位和细胞内pH值，并进一步恢复贴壁的人骨髓间充质干细胞的影响。我们的结果显示，与贴壁细胞和悬浮细胞相比，在1、5和10°C/min的冷却速率下冷冻保存后细胞活力显著降低约30%，表明贴壁细胞比悬浮细胞更脆弱。冷冻诱导的渗透压休克和冷休克导致细胞内性质的显著变化。10°C/min的冷却速率导致细胞内pH的酸化、丝状肌动蛋白的扭曲和积累以及线粒体的聚集。我们的研究结果还表明，1°C/min的冷却速率有助于维持细胞形态和丝状肌动蛋白的附着、完整性和均匀性，并导致冻存后更好的细胞恢复。
Cryopreservation is one of the most practical methods for the long-term storage of cell-matrix systems to ensure off-shelf availability in tissue engineering, stem cell therapy and drug testing. The aim of this study is to investigate the effects of osmotic and cold shock caused by the procedures of cryoprotectant agent addition/removal and freezing during cryopreservation on cell viability, intracellular properties, such as filamentous actin distribution, mitochondria localization and intracellular pH, and further recovery of adherent human mesenchymal stem cells. Our results shows a significant decrease in cell viability around 30% after cryopreservation at the cooling rates of 1, 5 and 10°C/min in comparison to the adherent cells and the cells in suspension, implicating that the adherent cells are more vulnerable than the suspension cells. The osmotic shock and cold shock induced by freezing lead to dramatic changes in the intracellular properties. The cooling rate of 10°C/min results in acidification of intracellular pH, distortion and accumulation of filamentous actin, and aggregation of mitochondria. Our findings also suggest that the cooling rate of 1°C/min helps to maintain cell morphology and attachment, integrity and uniformity of filamentous actin, and leads to better cell recovery after cryopreservation.