Department of Biochemistry, Institute for Problems of Cryobiology and Cryomedicine of the NAS Ukraine, Kharkiv, Ukraine; Department of Biomaterials and Tissue Engineering, Institute of Physiology of the CAS; Department of Neuroregeneration, Institute of Experimental Medicine of the CAS, Prague, Czech Republic,
Department of Biomaterials and Tissue Engineering, Institute of Physiology of the CAS; Department of Neuroregeneration, Institute of Experimental Medicine of the CAS, Prague, Czech Republic.
Cryo Letters. 2023 Mar-Apr;44(2):65-75.
Organoids represent indispensable opportunities for biomedicine, including drug discovery, cancer biology, regenerative and personalised medicine or tissue and organ transplantation. However, the lack of optimized preservation strategies limits the wide use of organoids in research or clinical fields. In this review, we present a short outline of the recent developments in organoid research and current cryopreservation strategies for organoid systems. While both vitrification and slow controlled freezing have been utilized for the cryopreservation of organoid structures or their precursor components, the controlled-rate slow freezing under protection of Me2SO remains the most common approach. The application of appropriate pre- or post-treatment strategies, like the addition of Rho-kinase or myosin inhibitors into cell culture or cryopreservation medium, can increase the recovery of complex organoid constructs post-thaw. However, the high complexity of the organoid structure and heterogeneity of cellular composition bring challenges associated with cryoprotectant distribution, distinct response of cells to the solution and freezing-induced injuries. The deficit of adequate quality control methods, which may ensure the assessment of organoid recovery in due term without prolonged re-cultivation process, represents another challenge limiting the reproducibility of current cryobanking technology. In this review, we attempt to assess the current demands and achievements in organoid cryopreservation and highlight the key questions to focus on during the development of organoid preservation technologies. Doi: 10.54680/fr23210110112.
类器官为生物医药提供了不可或缺的机会，包括药物发现、癌症生物学、再生和个性化医学或组织和器官移植。然而，缺乏优化的保存策略限制了类器官在研究或临床领域的广泛应用。在这篇综述中，我们简要介绍了类器官研究的最新进展和类器官系统的当前冷冻保存策略。虽然玻璃化和缓慢控制冷冻已被用于类器官结构或其前体细胞成分的冷冻保存，但在 Me2SO 保护下的控制速率缓慢冷冻仍然是最常见的方法。适当的预处理或后处理策略的应用，如在细胞培养或冷冻保存介质中添加 Rho-激酶或肌球蛋白抑制剂，可以增加解冻后复杂类器官结构的恢复。然而，类器官结构的高度复杂性和细胞组成的异质性带来了与冷冻保护剂分布、细胞对溶液的不同反应和冷冻诱导损伤相关的挑战。缺乏足够的质量控制方法是另一个限制当前冷冻库技术重现性的挑战，这些方法可以确保在不进行延长再培养过程的情况下及时评估类器官的恢复情况。在这篇综述中，我们试图评估类器官冷冻保存的当前需求和成就，并强调在开发类器官保存技术时需要关注的关键问题。doi: 10.54680/fr23210110112。
