Open AccessFeasibility of using cryopreserved lymphoblastoid cells to diagnose some lysosomal storage diseases
Author(s) -
De Mello A. S.,
Provin F.,
MichelinTireli K.,
Camelier M. V.,
Coelho J. C.
Publication year - 2010
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/j.1365-2184.2010.00660.x
Subject(s) - lymphoblast , mucopolysaccharidosis i , cell culture , cryopreservation , lysosomal storage disease , enzyme , biology , epstein–barr virus , cell , alpha galactosidase , immunohistochemistry , microbiology and biotechnology , virus , immunology , virology , biochemistry , fabry disease , pathology , enzyme replacement therapy , medicine , embryo , disease , genetics
Objectives: The Epstein–Barr virus (EBV) is utilized as a tool in the study of cellular biology because of its capacity to transform B‐lymphocytes. For this reason, EBV is used in conservation of human B‐lymphocytes for long periods for subsequent evaluation of lysosomal hydrolase activity . Lymphoblastoid cell lines have several advantages for use over other cell types, such as prompt availability and possibility to develop, characterize and standardize cell banks, to test effects of promising pharmaceutical reagents. The study below presents biochemical data that demonstrate validity of lymphoblastoid cell lines for diagnosis of GM1‐gangliosidosis, Gaucher, Fabry and Pompe diseases and mucopolysaccharidosis type I. Materials and methods: Cultures were prepared from peripheral blood, collected from 25 normal subjects and 13 affected individuals. Enzyme activities and immunohistochemistry (IHC) were measured. Activities of enzymes β‐galactosidase, β‐glucosidase, α‐iduronidase, α‐galactosidase and α‐glucosidase were measured before and after cryopreservation for 180 days. Enzymatic activity was measured when transformation was confirmed by IHC. Results: We observed some significant alterations in enzymatic activity of non‐cultured cells when compared to others that had been cultured for 12 days and kept frozen for 180 days. Conclusions: However, these alterations did not invalidate use of the technology of transformation of lymphoblastoid cell lines with EBV, to diagnose the diseases mentioned above, in view of the fact that the cultured cells, before and after freezing, demonstrated similar enzymatic activities.