Decot V, Houzé P, Stoltz J-F, Bensoussan D
CHU de Nancy, Cell Therapy and Tissues Unit, Brabois, Vandoeuvre-les-Nancy, France.
Biomed Mater Eng. 2009;19(4-5):293-300. doi: 10.3233/BME-2009-0594.
Dimethylsulfoxide (DMSO) is a cryoprotective substance often used to allow long term storage of stem cells or tissue grafts. However, a high frequency of adverse events is associated with the infusion of thawed cells. These events are in part due to DMSO, leading many cell therapy facilities to introduce a washing step before the delivery of the grafts. The lack of method for evaluating the residual quantities of this substance in the reinfused cells led us to develop a technique, based on capillary zone electrophoresis for assaying DMSO. The cryoprotectant was measured in 55 hematopoietic stem cell grafts, 6 parathyroids and 5 blood vessels immediately after thawing and after washing or bathing in a saline solution. The results showed that DMSO reduction in stem cell grafts reached more than 90% after the washing procedure. Furthermore, this study has shown that 2 washing steps significantly improved DMSO elimination as compared to 1 washing step. For parathyroids and blood vessels, bathing the tissues after thawing in a saline solution allowed more than 95% DMSO reduction. This study demonstrated that the technique of DMSO measurement used here, is simple and feasible on complex matrices such as protein samples after dilution. It is an appropriate method for residual quantification of the cryoprotectant before graft.
二甲基亚砜（DMSO）是一种冷冻保护剂，常用于长期保存干细胞或组织移植物。然而，解冻细胞输注会伴随高频率的不良事件。这些事件部分归因于DMSO，这使得许多细胞治疗机构在移植前增加了洗涤步骤。由于缺乏评估再输注细胞中该物质残留量的方法，我们开发了一种基于毛细管区带电泳的DMSO检测技术。在解冻后以及在盐溶液中洗涤或浸泡后，立即对55份造血干细胞移植物、6份甲状旁腺和5份血管样本中的冷冻保护剂进行了检测。结果显示，洗涤后干细胞移植物中的DMSO减少量超过90%。此外，该研究表明，与1次洗涤步骤相比，2次洗涤步骤能显著提高DMSO的清除率。对于甲状旁腺和血管，解冻后在盐溶液中浸泡可使DMSO减少量超过95%。本研究表明，此处使用的DMSO检测技术，对于稀释后的蛋白质样本等复杂基质简单可行。它是移植前冷冻保护剂残留量定量的合适方法。
