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

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.

Generation of Epstein-Barr Virus Immortalized-B Lymphocytes Cell Line of Orang Asli

“…LCL is not only capable to extend the proliferation to avoid the exhaustion of storage of precious DNA samples thus minimizes the need of resampling, but also it possesses identical genotypes and expression to their parental cells. Therefore it has been widely used as the resources in many biomedical studies, ranging from gene expression and functionality studies, proteomics and metabolomics, mutational analysis, to the population based genomic studies including Human Genome Diversity Project, HapMap and 1000 Genome Project [7][8][9][10][11][12][13][14][15][16][17][18][19][20][21].…”