Diffusion Limited Cryopreservation of Tissue with Radiofrequency Heated Metal Forms.
Diffusion Limited Cryopreservation of Tissue with Radiofrequency Heated Metal Forms.
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DOI：
10.1002/adhm.202000796
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发表时间：
2020-10
影响因子：
10
通讯作者：
Bischof JC
中科院分区：
文献类型：
--
作者：
Han Z;Sharma A;Gao Z;Carlson TW;O'Sullivan MG;Finger EB;Bischof JC
在生物医学应用中越来越需要冷冻保存的组织。然而，通常仅报告了薄组织（≤ 1 mm）的成功冷冻保存。这项工作提出了几项创新，以减少CPA毒性和改善组织冷冻保存，包括（1）通过射频金属形式和磁场优化提高组织升温速率，以及（2）实验验证的预测模型，以优化CPA加载和复温，以减少毒性。通过μCT成像研究CPA负载，通过热测量和建模进行复温，并在负载和/或冷冻保存后通过alamarBlue和组织学测量活力。设计了3种常见CPA混合物（6、8.4和9.3 M）的加载条件，然后快速冷却和金属形式复温（高达2000 °C/min）在具有各种CPA的冷冻保存的1-2 mm动脉中实现了≥ 90%的活力。尽管alamarBlue的存活率很高，但冷冻保存后组织学显示出细微的变化，表明细胞受到一定程度的损伤，特别是在厚达2 mm的动脉的中心部分。这些结果表明，小心的CPA加载和更高的金属形式升温速率可以帮助降低CPA加载毒性并改善组织中冷冻保存的结果，同时还提供了保存较大组织的新方案，厚度为1 mm。厚组织可以成功地冷冻保存和复温的帮助下，超快速金属形式加热。理论预测和实验实现了2 mm厚猪主动脉成功冻存复温的首例，并通过动脉组织验证了模型的有效性，该模型也可应用于其他组织系统。
Cryopreserved tissues are increasingly needed in biomedical applications. However, successful cryopreservation is generally only reported for thin tissues (≤ 1 mm). This work presents several innovations to reduce CPA toxicity and improve tissue cryopreservation, including (1) improved tissue warming rates through radiofrequency metal form and field optimization, and (2) an experimentally verified predictive model to optimize CPA loading and rewarming to reduce toxicity. CPA loading was studied by μCT imaging, rewarming by thermal measurements and modeling, and viability was measured after loading and/or cryopreservation by alamarBlue and histology. Loading conditions for 3 common CPA cocktails (6, 8.4 and 9.3 M) were designed and then fast cooling and metal forms rewarming (up to 2000 °C/min) achieved ≥ 90% viability in cryopreserved 1–2 mm arteries with various CPAs. Despite high viability by alamarBlue, histology showed subtle changes after cryopreservation suggesting some degree of cell damage especially in the central portions of thicker arteries up to 2 mm. While further studies are needed, these results show careful CPA loading and higher metal forms warming rates can help reduce CPA loading toxicity and improve outcomes from cryopreservation in tissues while also offering new protocols to preserve larger tissues ≥ 1 mm in thickness. Thick tissues can be successfully cryopreserved and rewarmed with the help of ultra-rapid metal form heating. The first case of successful cryopreservation and rewarming of 2 mm-thick porcine aorta is theoretically predicted and experimentally achieved, and the model which is validated by the arterial tissue can also be applied to other tissue systems.
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影响因子：
2.3
作者：
Fahy, Gregory M.;Wowk, Brian;Phan, Laura
通讯作者：
Phan, Laura
心脏瓣膜组织的玻璃化。
DOI：
10.1007/978-1-4939-2193-5_20
发表时间：
2015
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Brockbank KG;Chen Z;Greene ED;Campbell LH
通讯作者：
Campbell LH
影响因子：
2.7
作者：
BOUTRON, P
通讯作者：
BOUTRON, P
影响因子：
--
作者：
Etheridge, Michael L.;Xu, Yi;Bischof, John C.
通讯作者：
Bischof, John C.
影响因子：
2.2
作者：
Bastiaanse, J;Nanhekhan, LV;Egbrink, MGAO
通讯作者：
Egbrink, MGAO