In this concept article, we outline a variety of new approaches that have been conceived to address some of the remaining challenges for developing improved methods of biopreservation. This recognizes a true renaissance and variety of complimentary, high-potential approaches leveraging inspiration by nature, nanotechnology, the thermodynamics of pressure, and several other key fields. Development of an organ and tissue supply chain that can meet the healthcare demands of the 21st century means overcoming twin challenges of (1) having enough of these lifesaving resources and (2) having the means to store and transport them for a variety of applications. Each has distinct but overlapping logistical limitations affecting transplantation, regenerative medicine, and drug discovery, with challenges shared among major areas of biomedicine including tissue engineering, trauma care, transfusion medicine, and biomedical research. There are several approaches to biopreservation, the optimum choice of which is dictated by the nature and complexity of the tissue and the required length of storage. Short-term hypothermic storage at temperatures a few degrees above the freezing point has provided the basis for nearly all methods of preserving tissues and solid organs that, to date, have proved refractory to cryopreservation techniques successfully developed for single-cell systems. In essence, these short-term techniques have been based on designing solutions for cellular protection against the effects of warm and cold ischemia and basically rely upon the protective effects of reduced temperatures brought about by Arrhenius kinetics of chemical reactions. However, further optimization of such preservation strategies is now seen to be restricted. Long-term preservation calls for much lower temperatures and requires the tissue to withstand the rigors of heat and mass transfer during protocols designed to optimize cooling and warming in the presence of cryoprotective agents. It is now accepted that with current methods of cryopreservation, uncontrolled ice formation in structured tissues and organs at subzero temperatures is the single most critical factor that severely restricts the extent to which tissues can survive procedures involving freezing and thawing. In recent years, this major problem has been effectively circumvented in some tissues by using ice-free cryopreservation techniques based upon vitrification. Nevertheless, despite these promising advances there remain several recognized hurdles to be overcome before deep-subzero cryopreservation, either by classic freezing and thawing or by vitrification, can provide the much-needed means for biobanking complex tissues and organs for extended periods of weeks, months, or even years. In many cases, the approaches outlined here, including new underexplored paradigms of high-subzero preservation, are novel and inspired by mechanisms of freeze tolerance, or freeze avoidance, in nature. Others apply new bioengineering techniques such as nanotechnology, isochoric pressure preservation, and non-Newtonian fluids to circumvent currently intractable problems in cryopreservation.

中文翻译：

---

细胞、组织和器官冷冻保存的新方法

在这篇概念文章中，我们概述了各种新方法，这些新方法旨在解决开发改进的生物保存方法所面临的一些剩余挑战。这认可了真正的复兴和各种互补的、高潜力的方法，利用了自然、纳米技术、压力热力学和其他几个关键领域的灵感。开发能够满足 21 世纪医疗保健需求的器官和组织供应链意味着要克服双重挑战：(1) 拥有足够的救生资源；(2) 拥有存储和运输这些资源以用于各种应用的手段。每个都有独特但重叠的后勤限制，影响移植、再生医学和药物发现，生物医学主要领域面临共同的挑战，包括组织工程、创伤护理、输血医学和生物医学研究。生物保存有多种方法，最佳选择取决于组织的性质和复杂性以及所需的储存时间。在比冰点高几度的温度下进行短期低温储存，为几乎所有保存组织和实体器官的方法提供了基础，迄今为止，这些方法已被证明对为单细胞系统成功开发的低温保存技术是无效的。在本质上，这些短期技术​​的基础是设计针对细胞保护免受冷热缺血影响的解决方案，并且基本上依赖于化学反应的阿伦尼乌斯动力学带来的降低温度的保护作用。然而，目前看来，此类保存策略的进一步优化受到限制。长期保存需要低得多的温度，并要求组织在旨在优化冷冻保护剂存在下的冷却和升温的方案期间承受严酷的热和质量传递。现在人们普遍认为，采用目前的冷冻保存方法，在零度以下的温度下，结构化组织和器官中不受控制的冰形成是严重限制组织在涉及冷冻和解冻的过程中存活的最关键因素。近年来，通过使用基于玻璃化的无冰冷冻保存技术，一些组织中的这一主要问题已得到有效解决。然而，尽管取得了这些有希望的进展，但在通过经典冷冻和解冻或玻璃化进行深零度冷冻保存之前，仍然需要克服一些公认的障碍，才能为复杂组织和器官长时间保存数周、数月提供急需的手段。 ，甚至几年。在许多情况下，这里概述的方法，包括新的未经充分探索的高零度以下保存范例，是新颖的，并受到自然界中抗冻或避免冻结机制的启发。其他人则应用纳米技术、等容压保存和非牛顿流体等新的生物工程技术来解决目前冷冻保存中棘手的问题。