Fetal rhesus monkey lung cells can be grown in serum-free medium for the replication of dengue-2 vaccine virus |
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Authors: | Barbara Malewicz Lloyd E Anderson Karan Crilly Howard M Jenkin |
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Institution: | (1) The Hormel Institute, University of Minnesota, 801 16th Avenue, NE, 55912 Austin, Minnesota |
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Abstract: | Summary Serum-free media were developed to grow diploid fetal rhesus monkey lung (DBS-FRhL-2) cells and to propagate dengue-type 2
virus vaccine strain PR-159 (dengue-2 vaccine virus). Vitamins, amino acids, growth factors, hormones and other organic compounds,
and inorganic salts were substituted for fetal bovine serum. The composition of the medium that was optimal for growth of
DBS-FRhL-2 cells differed from medium optimal for the propagation of dengue-2 vaccine virus. Insulin, epidermal growth factor,
fibroblast growth factor, and platelet-derived growth factor were required for DBS-FRhL-2 cell proliferation in serum-free
medium but were inhibitory for virus propagation. Adenosine, cytidine, guanosine, uridine, and thymidine, each at 0.01 mM concentration, were necessary as medium supplements to obtain a high yield of dengue-2 vaccine virus in DBS-FRhL-2 cells
under serum-free conditions. DBS-FRhL-2 cells grown in serum-free medium produced dengue-2 vaccine virus with yields similar
to those of cells grown in the presence of serum. Dengue-2 vaccine virus obtained under serum-free conditions retained its
phenotypic markers such as temperature sensitivity and small plaque size.
This investigation was supported by Contract DAMD-17-81-C1029 from the U.S. Army Medical Research and Development Command,
and by The Hormel Foundation. |
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Keywords: | dengue virus vaccine serum-free media DBS-FRhL-2 cells growth factors plaque assay |
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