Tian Yunqing, Ma Yiming, Kang Yuxiang, Tian Shu, Li Qingsi, Zhang Lei, Yang Jing
Department of Biochemical Engineering, School of Chemical Engineering and Technology, Frontier Science Center for Synthetic Biology and Key Laboratory of Systems Bioengineering (MOE), Tianjin University, Tianjin, 300350, China.
Department of Minimally Invasive Spine Surgery, Tianjin Hospital, Tianjin University, Tianjin, 300211, China.
Acta Biomater. 2024 Sep 15;186:275-285. doi: 10.1016/j.actbio.2024.07.043. Epub 2024 Aug 2.
Hypothermic preservation (HP) is highly desired for the maintenance of the viability of living cell specimens, e.g. rare cells in whole-blood samples or therapeutic cells, in an unfrozen state. However, the extension of the viable preservation time is a challenge because of the multiple injuries suffered by hypothermically preserved cells. Here, based on a dynamic bond crosslinked zwitterionic hydrogel, we established a sensing preservation system that could monitor the levels of reactive oxygen species (ROS) via real-time electronic signals and intelligent control of antioxidant addition, to completely prevent an excess of ROS in the whole-cell specimen. Furthermore, the hydrogel-based system can counter the extracellular-matrix-loss-induced anoikis of living cells. Based on the design aimed at affording protection against two primary HP injuries (i.e. ROS overproduction and anoikis) to cells, this system extended the preservation time of cell specimens under refrigerated conditions to 24 days. After preservation, the use of a mild cell retrieval process guaranteed the activity of the preserved living cells. This work not only possesses the potential to facilitate intelligent cell-based clinical applications, but also paves the way for the preparation of living materials that can host programmed cells with long-term survival. STATEMENT OF SIGNIFICANCE: An intelligent system based on a zwitterionic sensing hydrogel is established, which can afford ultra-long hypothermic cell-preservation times of up to 24 days. The system enables the real-time monitoring of ROS overproduction and intelligent antioxidant addition, because of the merging of the smart hydrogel with a computer intelligent detection and control system. Furthermore, the automatic addition of an antioxidant according to the ROS-signal changes produced by the ZBA hydrogel effectively prevented HP lesions, including ROS over-production and ECM loss, in the preserved living cells. Subsequently, the system could also be gently dissociated, to retrieve the preserved cells. This work provides a solution for the real-time monitoring and long-term HP of living specimens, which holds the promise of benefiting cell-based medicine and the development of genetically programmed cell-based living materials.
低温保存（HP）对于维持活细胞标本（例如全血样本中的稀有细胞或治疗性细胞）在未冷冻状态下的活力非常必要。然而，由于低温保存的细胞会遭受多种损伤，延长其存活保存时间是一项挑战。在此，基于动态键交联两性离子水凝胶，我们建立了一种传感保存系统，该系统可以通过实时电子信号监测活性氧（ROS）水平并智能控制抗氧化剂的添加，以完全防止全细胞标本中ROS过量。此外，基于水凝胶的系统可以对抗细胞外基质丧失诱导的活细胞失巢凋亡。基于旨在为细胞提供针对两种主要HP损伤（即ROS过量产生和失巢凋亡）的保护的设计，该系统将细胞标本在冷藏条件下的保存时间延长至24天。保存后，使用温和的细胞回收过程可确保保存的活细胞的活性。这项工作不仅具有促进基于细胞的智能临床应用的潜力，而且还为制备能够长期存活地容纳程序化细胞的生物材料铺平了道路。重要性声明：建立了一种基于两性离子传感水凝胶的智能系统，该系统可提供长达24天的超长低温细胞保存时间。由于智能水凝胶与计算机智能检测和控制系统的融合，该系统能够实时监测ROS的过量产生并智能添加抗氧化剂。此外，根据ZBA水凝胶产生的ROS信号变化自动添加抗氧化剂，有效地防止了保存的活细胞中的HP损伤，包括ROS的过量产生和细胞外基质的丧失。随后，该系统也可以温和解离，以回收保存的细胞。这项工作为活标本的实时监测和长期HP提供了解决方案，有望造福基于细胞的医学和基因程序化细胞基生物材料的开发。
