Effect of Ice Nucleation and Cryoprotectants during High Subzero-Preservation in Endothelialized Microchannels,ACS Biomaterials Science & Engineering

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Effect of Ice Nucleation and Cryoprotectants during High Subzero-Preservation in Endothelialized Microchannels
ACS Biomaterials Science & Engineering ( IF 5.4 ) Pub Date : 2018-06-26 00:00:00 , DOI: 10.1021/acsbiomaterials.8b00648
Shannon N. Tessier _{1,

2,

3} , Lindong Weng _{1,

2} , Will D. Moyo _{1,

2} , Sam H. Au _{1,

2} , Keith H. K. Wong _{1,

2,

3} , Cindy Angpraseuth _{1,

2} , Amy E. Stoddard _{1,

2} , Chenyue Lu ₄ , Linda T. Nieman ₄ , Rebecca D. Sandlin _{1,

2,

3} , Korkut Uygun _{1,

2,

3} , Shannon L. Stott _{1,

4,

5} , Mehmet Toner _{1,

2,

3}

Affiliation

Cryopreservation is of significance in areas including tissue engineering, regenerative medicine, and organ transplantation. We investigated endothelial cell attachment and membrane integrity in a microvasculature model at high subzero temperatures in the presence of extracellular ice. The results show that in the presence of heterogeneous extracellular ice formation induced by ice nucleating bacteria, endothelial cells showed improved attachment at temperature minimums of −6 °C. However, as temperatures decreased below −6 °C, endothelial cells required additional cryoprotectants. The glucose analog, 3-O-methyl-d-glucose (3-OMG), rescued cell attachment optimally at 100 mM (cells/lane was 34, as compared to 36 for controls), while 2% and 5% polyethylene glycol (PEG) were equally effective at −10 °C (88% and 86.4% intact membranes). Finally, endothelialized microchannels were stored for 72 h at −10 °C in a preservation solution consisting of the University of Wisconsin (UW) solution, Snomax, 3-OMG, PEG, glycerol, and trehalose, whereby cell attachment was not significantly different from unfrozen controls, although membrane integrity was compromised. These findings enrich our knowledge about the direct impact of extracellular ice on endothelial cells. Specifically, we show that, by controlling the ice nucleation temperature and uniformity, we can preserve cell attachment and membrane integrity. Further, we demonstrate the strength of leveraging endothelialized microchannels to fuel discoveries in cryopreservation of thick tissues and solid organs.

中文翻译：

内皮化微通道中高度零度以下保存期间冰核和低温保护剂的作用

冷冻保存在组织工程，再生医学和器官移植等领域具有重要意义。我们研究了在存在细胞外冰的高零度以下温度下，微血管模型中的内皮细胞附着和膜完整性。结果表明，在由冰成核细菌诱导的异质细胞外冰形成的存在下，内皮细胞在最低-6°C的温度下显示出改善的附着性。但是，随着温度降低到-6°C以下，内皮细胞需要额外的冷冻保护剂。葡萄糖类似物3- O-甲基-d-葡萄糖（3-OMG），在100 mM时最佳挽救了细胞附着（细胞/泳道为34，对照组为36），而2％和5％的聚乙二醇（PEG）在-10°C时同样有效（ 88％和86.4％的完整膜）。最后，内皮化的微通道在-10°C的威斯康星大学（UW）溶液，Snomax，3-OMG，PEG，甘油和海藻糖组成的保存溶液中保存72 h。未冻结的控件，尽管膜的完整性受到了损害。这些发现丰富了我们关于细胞外冰对内皮细胞直接影响的知识。具体而言，我们表明，通过控制冰的成核温度和均匀度，我们可以保留细胞附着和膜的完整性。进一步，

更新日期：2018-06-26

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