移植保存液中脐带间充质干细胞活性与氧化应激的影响

Oxidative stress effect on viability of umbilical cord-derived mesenchymal stem cells in storage solution of transplantation

背景:目前临床移植常用的保存液可使人脐带来源间充质干细胞活性下降,影响移植效果,但关于其活性下降原因的报道目前还很少。目的:探索氧化应激是否为人脐带来源间充质干细胞临床移植保存过程中活性下降的因素;并在保存液中添加自由基清除剂是否可提高保存效果。方法:室温下用生理盐水保存人脐带来源间充质干细胞,0,2,4,6h后分别检测细胞内活性氧水平、检测丙二醛含量以及抗氧化酶活性以判定保存后细胞内氧化应激水平;保存液中添加自由基清除剂N-乙酰半胱氨酸后检测细胞贴壁率变化。结果与结论:经生理盐水保存后人脐带来源间充质干细胞内活性氧水平升高,细胞裂解液丙二醛含量呈时间依赖性增加,抗氧化酶超氧化物歧化酶、过氧化氢酶以及谷胱甘肽过氧化物酶活性均降低。添加N-乙酰半胱氨酸的移植保存液组比不添加组保存人脐带来源间充质干细胞后细胞活性氧水平降低、贴壁率升高。说明在临床常用移植保存液中人脐带来源间充质干细胞内活性氧升高是其活性下降的一个因素,在保存液中添加自由基清除剂N-乙酰半胱氨酸后能够一定程度的改善保存效果。

BACKGROUND： The viability of human umbilical cord-derived mesenchymal stem cells is often declined with the commonly used transplantation storage solution in clinics, which may influence the therapeutic effects of cellular transplantation. However, reasons for this are still unknown. OBJECTIVE： To investigate the role of oxidative stress in the reduction of human umbilical cord-derived mesenchymal stem cells viability in the storage process during clinical transplantation and to observe the effects of radical scavenger on the results. METHODS： Human umbilical cord-derived mesenchymal stem cells were harvested and cultured in normal saline for 0, 2, 4 and 6 hours at room temperature. Intracellular reactive oxygen levels were detected at those time points. Antioxidant enzyme activities and levels of malondialdehyde were measured to determine the intracellular oxidative stress levels after storage. Cell adhesion rate changes were retested after adding N-acetyl cysteine to the storage solution. RESULTS AND CONCLUSION： The reactive oxygen levels in human umbilical cord-derived mesenchymal stem cells were increased significantly after normal saline storage and levels of malondialdehyde were increased in a time-dependent manner. Activities of superoxide dismutase, catalase and glutathione peroxidase were all reduced. Addition of N-acetyl cysteine into the storage medium decreased the reactive oxygen levels and improved the human umbilical cord-derived mesenchymal stem cells viabilities. Experimental findings indicate that, increased reactive oxygen species in human umbilical cord-derived mesenchymal stem cells is one of the reasons for reduced cell viability. Adding the radical scavenger N-acetyl cysteine can improve the storage effects of human umbilical cord-derived mesenchymal stem cells.