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Observations on the growth requirements of vaccinia virus in vitro


, : Observations on the growth requirements of vaccinia virus in vitro. British Jour Exp Path 13(1): 90-96

The observation that vaccinia virus multiplies in a medium consisting of suitable minced tissue in Tyrode's solution without addition of serum, is confirmed. Virus grown in minced rabbit kidney and Tyrode's solution maintained its infectivity well when stored at [plus or minus] 5[degree]C. A proper ionic composition of the saline constituents of the medium is essential for growth of the virus. Tyrode's solution was suitable; buffered phosphate solution was not. Probably the tissue cells fail to survive in an unbalanced serum-phosphate mixture. Ready access of O to the tissue in the medium is essential for growth of the virus. It failed to grow when the tissue was covered by a layer of medium 15 mm. deep, and under these conditions respiration of the tissue rapidly ceased. The period for which minced rabbit kidney could be incubated at 37[degree] C. prior to inoculation, and still serve for cultivation of the virus, corresponded roughly with the period during which it respired. There ia a correlation between respiratory activity of tissues and their suitability to promote growth of virus; a correlation between the period a suitable tissue retained, during incubation in a saline medium at 37[degree]C. prior to inoculation, its property of promoting growth of the virus and the period for which it respired; and a correlation between the unsuitability of kidney in test-tubes under a layer of 15 mm. of medium and the absence of respiration. It was impossible to obtain cultures of virus in a cell-free kidney extract medium. Virus that had survived for 2-4 days in such a medium failed to grow when subcultured to a similar medium, but multiplied when transferred to a medium containing minced kidney. The exps. suggest that the condition maintained by suitable surviving cells, for cultivation of the virus, can not at present be otherwise obtained at will. In absence of fuller knowledge of the growth requirements of the virus, the significance of occasionally successful cultures in a cell-free medium remains obscure.

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