Cultivation of Vesicular Stomatitis Virus

Abstract

Carrel, Olitsky, and Long 1 reported the successful cultivation of the virus of vesicular stomatitis of horses in 1928. In their cultivation experiments, guinea pig tissues were used throughout. The method they employed consisted in placing embryonic tissue or adult bone marrow in contact with the virus which had been suspended in Tyrode's solution for 1 1/2 to 24 hours and then transferring to Carrel flasks containing coagulated plasma to which serum was added when marrow was used, or embryonic juice when embryonic tissue was used. The flasks were incubated for 7 to 10 days at 39°C.

No further reports of successful in vitro cultivation of the virus of vesicular stomatitis have been made.

This paper describes a much simplified method, similar to that employed by Li and Rivers^{2, 3} in the cutlivation of vaccine virus, for the cultivation of the virus of vesicular stomatitis. A satisfactory and readily available technique for the titration of the virus is given. Through the kindness of Dr. W. E. Cotton, samples were sent us of Indiana and New Jersey strains of virus which had been propagated in guinea pig pads for several years. The respective strains were then carried through 25 or more pad passages, filtered through Seitz filters, and this filtrate, or the Seitz filtrate of infected mouse brain tissue, 4 was used to initiate the cultures. The tissue employed was that of a 9 to 12 day chicken embryo which, after removal of the head and limbs, was minced in 3.5 cc. of Tyrode's solution. The Tyrode's solution was freshly prepared every 10 days according to the following formula: NaCl, 8.0 gm.; KCl, 0.2 gm.; CaCl₂, 0.2 gm.; MgCl₂, 0.1 gm.; NaH₂PO₄, 0.05 gm.; NaHCO₃, 0.5 gm.; glucose, 1.0 gm.; double distilled water q.s.ad. 1000.0 cc.; pH adjusted to 7.7 and sterilized by filtration through a Seitz or a tested Berkefeld N filter.