Steininger P A, Strasser E F, Ziehe B, Eckstein R, Rauh M
Department of Transfusion Medicine and Haemostaseology, University Hospital of Erlangen, Erlangen, Germany.
Department of Paediatrics and Adolescent Medicine, University Hospital of Erlangen, Erlangen, Germany.
Vox Sang. 2017 Feb;112(2):163-172. doi: 10.1111/vox.12482. Epub 2017 Jan 4.
Short-term storage of leukapheresis products used for immunotherapeutic mononuclear cell (MNC) products is a frequent event. The analysis of time-related metabolic patterns enables the characterization of storage-related effects in MNCs and the hypothesis-based optimization of the MNC medium.
The MNC products from seven leukapheresis procedures were stored within a closed bag system for 48 h. Concentrations of amino acids, biogenic amines, phospho- and sphingolipids and hexoses in the medium were measured by targeted metabolomics. The viability of MNC subpopulations was assayed by Annexin V (AnV) and JC-1 staining.
Glucose depletion and a significant change of the acylcarnitine profile are early events within the first 24 h of storage. In contrast, for most amino acids, the maximum increase was observed at 48 h of storage as mirrored by an increase in the amino acid levels by a mean factor of 1·2 (1·3, 2·0) after 6 h (24 h, 48 h, respectively). This was except for the concentrations of glutamine and lysine, which did not change significantly. The taurine concentration showed a twofold increase within the first 24 h and remained constant thereafter. The steepest increase in AnV and 7-AAD CD4 T cells was found between 24 and 48 h.
The time-course of apoptosis and metabolic patterns in the MNC products demonstrate that 24 h of storage is a decisive time-point, as afterwards key metabolic pathways showed nonlinear detrimental changes. Optimization of storage by supplementation of specific substrates demands therefore an early intervention.
用于免疫治疗的单核细胞（MNC）产品的白细胞分离术产品的短期储存是常有的事。对与时间相关的代谢模式进行分析，能够对MNC中与储存相关的效应进行表征，并基于假设对MNC培养基进行优化。
将来自7次白细胞分离术程序的MNC产品在封闭袋系统中储存48小时。通过靶向代谢组学测量培养基中氨基酸、生物胺、磷酸和鞘脂以及己糖的浓度。通过膜联蛋白V（AnV）和JC-1染色测定MNC亚群的活力。
葡萄糖消耗和酰基肉碱谱的显著变化是储存前24小时内的早期事件。相比之下，对于大多数氨基酸，在储存48小时时观察到最大增加，6小时（分别为24小时、48小时）后氨基酸水平平均增加1.2倍（1.3、2.0）反映了这一点。谷氨酰胺和赖氨酸的浓度除外，它们没有显著变化。牛磺酸浓度在最初24小时内增加了两倍，此后保持不变。在24至48小时之间发现AnV和7-AAD CD4 T细胞的增加最为明显。
MNC产品中细胞凋亡和代谢模式的时间进程表明，储存24小时是一个决定性的时间点，因为此后关键代谢途径显示出非线性有害变化。因此，通过补充特定底物来优化储存需要早期干预。
