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首页> 外文期刊>Biopreservation and biobanking >Cryopreservation of Porcine Aortic Heart Valve Leaflet-Derived Myofibroblasts
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Cryopreservation of Porcine Aortic Heart Valve Leaflet-Derived Myofibroblasts

机译：猪主动脉心脏瓣膜小叶成肌纤维细胞的冷冻保存

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Cryopreservation by freezing is usually employed for storage of allogeneic valves destined for clinical use; however, disruption of leaflet extracellular matrices by ice may occur. This study was performed to determine the effects of cryoprotectants employed in VS55 for ice-free, vitreous cryopreservation upon adherent porcine heart valve leaflet-derived myofibroblasts in culture. Low-passage myofibroblasts exhibiting strong actin and myosin staining were employed for these experiments. Three cryoprotectants, dimethylsulfoxide, formamide, and propanediol, were tested individually and in combination. Exposure experiments demonstrated that the individual cryoprotectants (0-5 M) were generally cytotoxic in a dose-dependent manner with little if any loss of cells as determined by measuring metabolic activity and DNA content. Exposure to formamide resulted in the greatest loss of cells and reduction in viability. Combination of the three cryoprotectants demonstrated that the cytotoxic effects of each cryoprotectant were not cumulative. Cell viability and DNA content were equivalent to dimethylsulfoxide and propanediol and higher than formamide alone over most of the 0-5 M dose—response curve. After cryopreservation by slow-rate freezing, the benefits of the combination of cryoprotectants over individual cryoprotectants were demonstrated at the 4 M concentration range for both cell viability and cell retention. In conclusion, these studies demonstrated that, with the exception of the lower concentrations of propanediol, the combination of cryoprotectants employed in these studies result in equivalent or better cell viability and attachment than individual cryoprotectants.
机译：通常通过冷冻冷冻保存用于临床用途的同种异体瓣膜。然而，冰可能会破坏小叶细胞外基质。进行这项研究是为了确定VS55中用于无冰，玻璃体冷冻保存的冷冻保护剂对培养中粘附猪心瓣小叶的成纤维细胞的影响。这些实验采用表现出强烈的肌动蛋白和肌球蛋白染色的低通道成肌纤维细胞。分别或组合测试了三种防冻剂，即二甲基亚砜，甲酰胺和丙二醇。暴露实验表明，单个的冷冻保护剂（0-5 M）通常具有剂量依赖性的细胞毒性，通过测量代谢活性和DNA含量可以确定几乎没有细胞损失。暴露于甲酰胺会导致最大的细胞损失和活力降低。三种冷冻保护剂的组合表明每种冷冻保护剂的细胞毒性作用不是累积的。在0-5 M的大部分剂量反应曲线中，细胞活力和DNA含量相当于二甲基亚砜和丙二醇，高于单独的甲酰胺。在通过慢速冷冻进行冷冻保存后，在4 M的浓度范围内，无论是细胞存活力还是细胞保留率，冷冻保护剂组合相对于单个冷冻保护剂的益处都得到了证明。总之，这些研究表明，除了较低浓度的丙二醇外，这些研究中使用的冷冻保护剂组合可产生比单个冷冻保护剂相同或更好的细胞活力和附着力。

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