Vitreous Cryopreservation of Human Umbilical Vein Endothelial Cells with Low Concentration of Cryoprotective Agents for Vascular Tissue Engineering.
Vitreous Cryopreservation of Human Umbilical Vein Endothelial Cells with Low Concentration of Cryoprotective Agents for Vascular Tissue Engineering.
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DOI：
10.1089/ten.tec.2016.0335
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发表时间：
2016-09
期刊：
影响因子：
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通讯作者：
Yu-xuan Zheng;Gang Zhao;F. Panhwar;Xiaoming He
中科院分区：
文献类型：
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作者：
Yu-xuan Zheng;Gang Zhao;F. Panhwar;Xiaoming He
人脐静脉内皮细胞（HUVEC）的冷冻保存对于组织工程应用以及内皮细胞在心脑血管疾病中的作用研究具有重要意义。使用高浓度冷冻保护剂 (CPA) 和缓慢冷冻的传统玻璃化冷冻方法（将样品冷却到低温但没有明显冻结）和缓慢冷冻分别由于高浓度 CPA 的严重毒性和冰形成引起的低温损伤而不是最佳方法。在这项研究中，我们开发了一种通过低浓度 CPA 玻璃化冷冻保存 HUVEC 的方法。这是通过优化 CPA 并使用高导热石英毛细管 (QC) 来容纳玻璃化样品来实现的。后者最大限度地减少热质量以产生超快的冷却/升温速率。我们的数据表明，使用 1.4 mol/L 乙二醇和 1.1 mol/L 二甲基亚砜可以在 QC 中玻璃化 HUVEC，且存活率超过 90%。此外，该方法显着提高了冷冻保存的 HUVEC 的附着效率。冷冻保存后贴壁细胞的增殖与新鲜细胞类似。因此，这项研究可能提供一种有效的玻璃化技术来储存 HUVEC，用于血管组织工程和其他应用。
Cryopreservation of human umbilical vein endothelial cells (HUVECs) is important to tissue engineering applications and the study of the role of endothelial cells in cardiovascular and cerebrovascular diseases. The traditional methods for cryopreservation by vitrification (cooling samples to a cryogenic temperature without apparent freezing) using high concentration of cryoprotective agents (CPAs) and slow freezing are suboptimal due to the severe toxicity of high concentration of CPAs and ice formation-induced cryoinjuries, respectively. In this study, we developed a method to cryopreserve HUVECs by vitrification with low concentration of CPAs. This is achieved by optimizing the CPAs and using highly thermally conductive quartz capillary (QC) to contain samples for vitrification. The latter minimizes the thermal mass to create ultra-fast cooling/warming rates. Our data demonstrate that HUVECs can be vitrified in the QC using 1.4 mol/L ethylene glycol and 1.1 mol/L dimethyl sulfoxide with more than 90% viability. Moreover, this method significantly improves the attachment efficiency of the cryopreserved HUVECs. The attached cells post-cryopreservation proliferate similarly to fresh cells. Therefore, this study may provide an effective vitrification technique to bank HUVECs for vascular tissue engineering and other applications.