一年的冷冻保存对EBV转化淋巴细胞溶酶体水解酶活性的影响。
Effect of one year of cryopreservation on the activity of lysosomal hydrolases from EBV-transformed lymphocytes.
作者信息
de Mello A S, Mendes F B, Michelin-Tireli K, Camelier M V, Coelho J C
机构信息
Postgraduate Program in Biological Sciences and Biochemistry, Department of Biochemistry, Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS, Brazil.
出版信息
J Biomed Biotechnol. 2011;2011:132581. doi: 10.1155/2011/132581. Epub 2011 May 19.
BACKGROUND
The Epstein-Barr virus (EBV) was used as an agent of B lymphocyte proliferation for subsequent diagnosis of lysosomal storage disease. Due to the constant handling of long-preserved samples in our cell bank, we decided to observe the behavior and then compare cultured and frozen samples for at least one year's cryopreservation.
METHODS
Twenty-five samples from healthy individuals were used to assess the possible changes in activity of enzymes β-galactosidase, β-glucosidase, α-iduronidase, α-galactosidase, and α-glucosidase. Transmission electron microscopy was used to confirm cell transformation of B lymphocytes into EBV-infected cells, generating lymphoblastoid cell lines.
RESULTS
Transmission electron microscopy findings confirmed previous reports in the literature that is, significant and evident morphological changes in the nucleus occur after day 12 and the consequent cell transformation into EBV-infected cells. After thawing and subsequent treatment with the five enzymes utilized, we observed no significant changes in samples cryopreserved for more than one year, as compared to samples cultured for 12 days.
背景
爱泼斯坦-巴尔病毒（EBV）被用作B淋巴细胞增殖的因子，用于随后溶酶体贮积病的诊断。由于我们细胞库中长期保存样本的持续处理，我们决定观察其行为，然后比较培养样本和冷冻样本至少一年的冷冻保存情况。
方法
使用来自健康个体的25个样本评估β-半乳糖苷酶、β-葡萄糖苷酶、α-L-艾杜糖苷酶、α-半乳糖苷酶和α-葡萄糖苷酶活性的可能变化。透射电子显微镜用于确认B淋巴细胞向EBV感染细胞的细胞转化，从而生成淋巴母细胞系。
结果
透射电子显微镜检查结果证实了文献中先前的报道，即第12天后细胞核出现显著且明显的形态变化，随后细胞转化为EBV感染细胞。在解冻并随后用所使用的五种酶处理后，与培养12天的样本相比，我们观察到冷冻保存一年以上的样本没有显著变化。
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