Method of Suspension Culture for MDCK Cells and Isolation of Influenza Virus in MDCK Suspension Cultured Cells

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  • MDCK細胞の浮遊培養および浮遊培養細胞を用いたインフルエンザウイルスの分離について
  • MDCK サイボウ ノ フユウ バイヨウ オヨビ フユウ バイヨウ サイボウ

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Abstract

Studies were made of a method of suspension culture for MDCK cells which enables us to prepare a sufficient quantity of cells in a regularly available condition in preparation for an unexpected occurrence of influenza epidemics, and examination of HA yield in suspension cultured cells and virus isolation from patient's specimen were performed. A silicon coated spinner flask as culture bottle and a Joklik modification of MEM added with foetal bovine serum, tryptose phosphate broth and methyl cellulose solutions as growth medium. Examination of HA yield with the inoculation 1.5×103PFU/ml of viruses (average quantity of viruses in throat washings) upon the suspension cultured cells in 8 steps from 1.1×106 cells/ml to 9.2×103 cells/ml of cells revealed that HA yield became better in higher concentration of cells to 2.3×105 cells/ml, but conversely fell down in much higher concentration. In contrast with HA yield with the monolayer cells, HA yield with the suspension cultured cells was poor with low m.o.i.<BR>Using 2×105 cells/ml of suspension cultured cells, 20 strains of influenza virus were isolated from 60 specimens. The rate of isolation was a little lower than that with the fertile hen's egg (27 strains were isolated), but the number of days necessary for the pathogenic determination could be lessend.

Journal

  • Kansenshogaku Zasshi

    Kansenshogaku Zasshi 54 (6), 306-312, 1980

    The Japanese Association for Infectious Diseases

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