Guidance for Removal of Fetal Bovine Serum from Cryopreserved Heart Valve Processing
Guidance for Removal of Fetal Bovine Serum from Cryopreserved Heart Valve Processing
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DOI：
10.1159/000321166
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发表时间：
2011-01-01
影响因子：
2.7
通讯作者：
Schenke-Layland, Katja
中科院分区：
文献类型：
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作者：
Brockbank, Kelvin G. M.;Heacox, Albert E.;Schenke-Layland, Katja
牛血清通常用于同种异体心脏瓣膜的冷冻保存;然而，牛血清存在因牛源性感染因子污染而导致产品掺假的风险。在本研究中，我们比较了新鲜和冷冻保存的猪瓣膜，这些瓣膜采用4种冷冻保存制剂中的1种进行处理，其中3种无血清，1种使用含1.4 M二甲亚砜的牛血清。在第一个无血清组中，简单地从冷冻保存制剂中去除牛血清。第二种无血清制剂具有更高的冷冻保护剂浓度，即2 M二甲亚砜，与无血清溶液组合。由于理论上考虑到去除血清可能会增加组织破裂的发生率，将胶体葡聚糖40添加到第三个无血清组中，其中2添加到二甲基亚砜中。复温后，检查瓣膜并进行一系列试验。大体病理学显示98个冷冻心脏瓣膜中有1个导管破裂。冷冻保存组相对于新鲜组的活力数据表明一半的比较中活力丧失（p < 0.05）。在有或没有牛血清的任何冷冻保存组之间均未观察到显著差异。无论是常规组织学，自发荧光为基础的多光子成像，也没有半定量二次谐波显微镜细胞外基质成分显示任何统计学上的显着差异。生物力学分析也显示无显著差异。我们的结果表明，牛血清可以安全地从心脏瓣膜加工中去除，并且不需要胶体来防止破裂。本研究为评估组织冷冻保存程序的变化提供了指导。版权所有（C）2010 S. Karger AG，巴塞尔
Bovine serum is commonly used in cryopreservation of allogeneic heart valves; however, bovine serum carries a risk of product adulteration by contamination with bovine-derived infectious agents. In this study, we compared fresh and cryopreserved porcine valves that were processed by 1 of 4 cryopreservation formulations, 3 of which were serum-free and 1 that utilized bovine serum with 1.4 M dimethylsulfoxide. In the first serum-free group, bovine serum was simply removed from the cryopreservation formulation. The second serum-free formulation had a higher cryoprotectant concentration, i.e. 2 M dimethylsulfoxide, in combination with a serum-free solution. A colloid, dextran 40, was added to the third serum-free group with 2 to dimethylsulfoxide due to theoretical concerns that removal of serum might increase the incidence of tissue cracking. Upon rewarming, the valves were inspected and subjected to a battery of tests. Gross pathology revealed conduit cracking in 1 of 98 frozen heart valves. Viability data for the cryopreserved groups versus the fresh group demonstrated a loss of viability in half of the comparisons (p < 0.05). No significant differences were observed between any of the cryopreserved groups, with or without bovine serum. Neither routine histology, autofluorescence-based multiphoton imaging nor semiquantitative second-harmonic generation microscopy of extracellular matrix components revealed any statistically significant differences. Biomechanics analyses also revealed no significant differences. Our results demonstrate that bovine serum can be safely removed from heart valve processing and that a colloid to prevent cracking was not required. This study provides guidance for the assessment of changes in cryopreservation procedures for tissues. Copyright (C) 2010 S. Karger AG, Basel