Propagation of Rabies Virus in Tissue Culture.

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Abstract
The rabies virus was propagated for as long as 57 passages (transfers) in cultures of chopped mouse embryo brain suspended in rabbit serum and buffered salt solns. Under conditions developed in shorter expts., the virus multiplied to the extent that mice were killed regularly by the culture material in dilutions as high as 10-6 (1:1,000,000); and it was possible to produce typical rabies in mice with 1/33,000,000 ml. of the culture virus. When ox serum ultra-filtrate was substituted for rabbit serum, the virus titer was somewhat less. Comparative expts. showed the importance of grinding the culture material for mouse titra-tions. No significant increase in virus output was ever obtained by subjecting the cultures to continuous gassing with mixtures of O2, CO2 and N2. A duplicate series of cultures incubated for 9 passages at 35[degree] and at 37.5[degree] C, respectively, showed no consistent difference in the amt. of virus produced. Attempts to cultivate the rabies virus in chick embryo brain cultures were unsuccessful even when the virus had been adapted to chick tissue by serial intra-cerebral passage in embryonated eggs. Rabies virus from a mouse brain frozen at [long dash]60[degree] C for 60 days was cultivated for 24 successive passages in cultures of brain tissue from embryo mice (5 passages), 1-day-old mice (3 passages), 3-day-old mice (2 passages), 5-day-old mice (10 passages) and 14-day-old mice (4 passages), respectively. Culture virus developed in brain material from 5-day-old mice failed to multiply in cultures of brain tissue from 48-day-old mice.

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