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Journal of Biomedicine and Biotechnology
Volume 2011, Article ID 132581, 5 pages
http://dx.doi.org/10.1155/2011/132581
Research Article

Effect of One Year of Cryopreservation on the Activity of Lysosomal Hydrolases from EBV-Transformed Lymphocytes

1Postgraduate Program in Biological Sciences and Biochemistry, Department of Biochemistry, Federal University of Rio Grande do Sul, 90035-003 Porto Alegre, RS, Brazil
2Medical Genetics Service, Clinical Hospital of Porto Alegre, 90035-003 Porto Alegre, RS, Brazil

Received 25 November 2010; Revised 15 March 2011; Accepted 23 March 2011

Academic Editor: Ayman El-Kadi

Copyright © 2011 A. S. de Mello et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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.