Use of serum substitutes for the growth of four cell lines commonly used for virus isolation.
Four serum extenders (Opti-MEM I, BioRich, Excyte I and Excyte III) and two serum substitutes (Omni Serum and Serum Plus) were evaluated for the reduction or replacement of FBS for the growth and maintenance of four representative cell lines used for virus isolation. MRC-5, human fetal foreskin fibroblast (HF), A549 and Hep-2 cells were seeded into culture flasks containing MEM with 10% FBS or with the serum extenders or substitutes and subcultured into 16 x 125 mm glass tubes and 1-dram shell vials. Only cells cultured with Opti-MEM I and Omni Serum grew consistently in tubes and vials and these reagents were compared to FBS for viral isolation and detection. Laboratory stocks of CMV, HSV, VZV, adenovirus (Ad) and RSV were tested. In HF and MRC-5 cells, CMV was isolated in cells cultured in either Opti-MEM I or Omni Serum before CPE appeared in cultures containing FBS. Ad and VZV CPE were observed first in HF and MRC-5 cells containing Omni Serum. HSV CPE was seen at the same time in HF and MRC-5 cells with all 3 media. HSV and Ad CPE in A549 cells were also seen at the same time in all 3 media. RSV and Ad CPE in Hep-2 cells were observed first in FBS containing media. Both Opti-MEM I and Omni Serum performed well as substitutes for FBS for growth of stock viruses without loss of cell sensitivity.
Gleaves
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Diagnostic Virology Laboratory, Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
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Meyers
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D000818:Animals; D001802:Blood Substitutes; D002417:Cattle; D002460:Cell Line; D002478:Cells, Cultured; D003470:Culture Media; D004267:DNA Viruses / Q000254:growth & development / Q000302:isolation & purification; D006801:Humans; D012136:Respiratory Syncytial Viruses / Q000254:growth & development / Q000302:isolation & purification; D014776:Virus Cultivation
D001802:Blood Substitutes; D003470:Culture Media