{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T04:46:17Z","timestamp":1790916377610,"version":"4.1.0"},"reference-count":19,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2006,1,19]],"date-time":"2006-01-19T00:00:00Z","timestamp":1137628800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cell Proliferation"],"published-print":{"date-parts":[[2006,2]]},"abstract":"<jats:p><jats:bold>Abstract.\u2002<\/jats:bold> Epstein\u2013Barr virus (EBV) infection <jats:italic>in vitro<\/jats:italic> causes transformation of B cells and generates B lymphoblastoid cell lines (LCLs). These LCLs have been widely used for the diagnostic of several genetic metabolic disorders. However, up to now, efficiency of LCL generation has been based on misleading subjective analysis. In this study, quantitative analyses have been performed to indicate efficiency of B\u2010cell transformation to measuring human lysosomal acid hydrolases associated with: GM1\u2010gangliosidosis type I, Gaucher disease and mucopolysaccharidosis type I. Peripheral blood mononuclear cells were isolated from 13 subjects, and LCLs were produced by culturing them with EBV for 12\u00a0days. Activities of the enzymes \u03b2\u2010galactosidase, \u03b2\u2010glucosidase and \u03b1\u2010iduronidase were measured before and after cryopreservation in liquid nitrogen for 30\u00a0days. Efficiency of the B\u2010cell transformation was screened every 4\u00a0days by the enumeration of cell proliferation, cell counts and changes in granularity estimated by flow cytometry. We observed the generation of 13 LCLs. Cell transformation was confirmed by the gradual increase of cellular clusters, cell size and granularity. In addition, we determined that the activity of the enzymes mentioned above did not change following cryopreservation. These data suggest that our enumerative approach for screening of EBV\u2013LCLs is efficient for the enzymatic determination of human lysosomal acid hydrolases and may thus replace misleading subjective analyses.<\/jats:p>","DOI":"10.1111\/j.1365-2184.2006.00363.x","type":"journal-article","created":{"date-parts":[[2006,1,19]],"date-time":"2006-01-19T17:47:28Z","timestamp":1137692848000},"page":"29-36","source":"Crossref","is-referenced-by-count":9,"title":["Epstein\u2013Barr virus\u2010induced transformation of B cells for the diagnosis of genetic metabolic disorders \u2013 enumerative conditions for cryopreservation"],"prefix":"10.1111","volume":"39","author":[{"given":"A. 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