低温保存(HP)是非常需要的活细胞标本的生存能力的维持,例如全血样本中的稀有细胞或治疗细胞,处于未冻结状态。然而,由于低温保存的细胞遭受多重伤害,延长存活保存时间是一个挑战。这里,基于动态键交联两性离子水凝胶,我们建立了一个传感保存系统,可以通过实时电子信号和抗氧化剂添加的智能控制来监测活性氧(ROS)的水平,以完全防止全细胞标本中过量的ROS。此外,基于水凝胶的系统可以对抗细胞外基质损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过量生产和失巢)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保证了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,但也为制备能够长期存活的程序化细胞的活材料铺平了道路。重要声明:建立了基于两性离子传感水凝胶的智能系统,这可以提供长达24天的超长低温细胞保存时间。该系统实现了对ROS生产过剩和智能抗氧化剂添加的实时监测,因为智能水凝胶与计算机智能检测和控制系统的合并。此外,根据ZBA水凝胶产生的ROS信号变化自动添加抗氧化剂可有效预防HP病变,包括ROS超量生产和ECM损失,保存在活细胞中。随后,该系统也可以轻轻解离,检索保存的细胞。这项工作为活体标本的实时监测和长期HP提供了解决方案,它有望使基于细胞的医学和遗传编程的基于细胞的活材料的开发受益。
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.