Gangwar Lakshya, Han Zonghu, Scheithauer Cameron, Namsrai Bat-Erdene, Kantesaria Saurin, Goldstein Rob, Etheridge Michael L, Finger Erik B, Bischof John C
Department of Mechanical Engineering, University of Minnesota, Minneapolis, USA.
Department of Surgery, University of Minnesota, Minneapolis, USA.
bioRxiv. 2024 Nov 11:2024.11.08.622572. doi: 10.1101/2024.11.08.622572.
Organ banking by vitrification could revolutionize transplant medicine. However, vitrification and rewarming have never been demonstrated at the human organ scale. Using modeling and experimentation, we tested the ability to vitrify and rewarm 0.5 - 3 L volumes of three common cryoprotective agent (CPA) solutions: M22, VS55, and 40% EG+0.6M Sucrose. We first demonstrated our ability to avoid ice formation by convectively cooling faster than the critical cooling rates of these CPAs while also maintaining adequate uniformity to avoid cracking. Vitrification success was then verified by visual, thermometry, and x-ray μCT inspection. M22 and EG+sucrose were successfully vitrified in 0.5 L bags, but only M22 was vitrified at 3 L. VS55 did not vitrify at any tested volumes. As additional proof of principle, we successfully vitrified a porcine liver (~1L) after perfusion loading with 40% EG+0.6M Sucrose. Uniform volumetric rewarming was then achieved in up to 2 L volumes (M22 with ~5 mgFe/mL iron-oxide nanoparticles) using nanowarming, reaching a rate of ~88 °C/min with a newly developed 120 kW radiofrequency (RF) coil operating at 35kA/m and 360kHz. This work demonstrates that human organ scale vitrification and rewarming is physically achievable, thereby contributing to technology that enables human organ banking.
通过玻璃化进行器官保存可能会彻底改变移植医学。然而,玻璃化和复温从未在人体器官尺度上得到证实。我们通过建模和实验,测试了对三种常见冷冻保护剂(CPA)溶液(M22、VS55和40%乙二醇+0.6M蔗糖)0.5 - 3升体积进行玻璃化和复温的能力。我们首先证明了通过对流冷却速度快于这些CPA的临界冷却速率来避免结冰的能力,同时还保持了足够的均匀性以避免破裂。然后通过视觉、温度测量和X射线μCT检查验证了玻璃化的成功。M22和乙二醇+蔗糖在0.5升袋子中成功实现了玻璃化,但只有M22在3升时实现了玻璃化。VS55在任何测试体积下都未实现玻璃化。作为原理的进一步证明,我们在用40%乙二醇+0.6M蔗糖灌注加载后成功地对一个猪肝(约1升)进行了玻璃化。然后使用纳米复温在高达2升体积(含有约5毫克铁/毫升氧化铁纳米颗粒的M22)中实现了均匀的体积复温,使用新开发的在35kA/m和360kHz下运行的120千瓦射频(RF)线圈,复温速率达到约88℃/分钟。这项工作表明人体器官尺度的玻璃化和复温在物理上是可以实现的,从而为实现人体器官保存的技术做出了贡献。
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