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Interaction of the LM cell surface with methylcellulose and vaccinia virus

Mode of action and implications for large scale vaccine production

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Summary

The adsorption of methylcellulose (MC) by LM mouse cells from phosphate-buffered saline (PBS), Hanks' solution without glucose (HBSS), and medium 199 (199) with or without peptone (P) has been demonstrated by chemical and radioactive methods. A fixed amount of MC was removed from PBS and HBSS containing MC, whereas cells suspended in 199P containing MC were capable of taking up larger amounts as the MC concentration increased. Suspension and monolayer cells behaved similarly. Treatment with 2,4-dinitrophenol did not inhibit adsorption in PBS. The osmotic pressure, viscosity, and surface tension for these solutions indicated that physical forces were involved in the effects of MC on cells. MC at low concentrations reacted with mono- and divalent cations. Autoradiography showed the presence of MC at or near the surface of cells. Vaccinia virus uptake by monolayer cells was delayed but not inhibited. This appeared to be related to the calculated diffusion coefficients of the virus. Magnesium ions reduced the plaque titer in a manner independent of the MC effect. Electron microscopy demonstrated the adherence of several virus particles per cell at the membrane or on the microvilli, with or without MC treatment, but cell clumping occurred more readily without MC. It is concluded that several chemical and physical effects account for the behavior of LM cells in MC-containing media. Use of MC thus makes possible large scale virus production in LM cells in protein-free medium.

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Author notes

  1. Donald J. Merchant

    Present address: W. Alton Jones Cell Science Center, P. O. Box 631, 12946, Lake Placid, N. Y.

Authors and Affiliations

  1. Department of Bacteriology and Immunology, Health Sciences Centre, University of Western Ontario, 72, London, Ontario, Canada

    Edward L. Medzon

  2. Department of Microbiology, University of Michigan, 48104, Ann Arbor, Michigan

    Donald J. Merchant

Authors
  1. Edward L. Medzon
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Medzon, E.L., Merchant, D.J. Interaction of the LM cell surface with methylcellulose and vaccinia virus. In Vitro 7, 46–58 (1971). https://doi.org/10.1007/BF02619004

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