Xu Rui, Brookshaw Tom, Erro Eloy, Selden Clare, Martin Eleanor
Department of Medical Physics and Biomedical Engineering, University College London, London, UK.
Wellcome / EPSRC Centre for Interventional and Surgical Sciences, University College London, London, UK.
Sci Rep. 2025 Oct 28;15(1):37664. doi: 10.1038/s41598-025-21464-x.
Rapid volumetric rewarming methods are needed to enable the effective cryopreservation and recovery of large volumes of biological cells for therapy and banking of tissues and organs. Ultrasonic rewarming is currently under development, but its effect on cells and their post-rewarming viability has not yet been established. Here, we compare ultrasonic rewarming with the gold-standard [Formula: see text]C water bath using cryovials containing cryopreserved alginate encapsulated liver spheroids. Mean rewarming rates are used to establish the exposure time to rewarm to [Formula: see text]C for higher power (100 W) and lower power (20 W) ultrasonic rewarming. These electrical powers correspond to free-field pressures along the central cryovial axis of 2.8 MPa and 1.3 MPa, respectively. Ultrasonic rewarming is faster than the gold-standard (120±5 s), taking 88 s (36% faster) and 34 s (350% faster) to rewarm to [Formula: see text]C with the lower and higher powers. We measure post-rewarming liver spheroid viability and viable cell number across the 96-h recovery period. The lower power improves viability by 1% and the higher power reduces viability by 2% on average, relative to the gold-standard. There were no significant differences in viable cell number between rewarming methods. Our findings will serve as a foundation for ultrasonic cryovial rewarming and demonstrates potential for scaling to larger volumes.
需要快速的体积复温方法，以实现对大量生物细胞进行有效的冷冻保存和复苏，用于组织和器官的治疗及储存。超声复温目前正在研发中，但其对细胞及其复温后活力的影响尚未确定。在此，我们使用装有冷冻保存的藻酸盐包封肝球的冻存管，将超声复温与金标准37℃水浴进行比较。使用平均复温速率来确定在较高功率（100W）和较低功率（20W）超声复温下复温至37℃的暴露时间。这些电功率分别对应沿冻存管中心轴的自由场压力为2.8MPa和1.3MPa。超声复温比金标准（120±5秒）更快，在较低功率和较高功率下复温至37℃分别需要88秒（快36%）和34秒（快350%）。我们在96小时的恢复期内测量复温后肝球的活力和活细胞数量。相对于金标准，较低功率平均使活力提高1%，较高功率平均使活力降低2%。复温方法之间的活细胞数量没有显著差异。我们的研究结果将为超声冻存管复温奠定基础，并证明了扩大到更大体积的潜力。
