Viruses and other pathogens can diverge in their evolved host-use strategies, due to exposure to different host types and conflicts between within-host reproduction and between-host survival. Most host-pathogen studies have emphasized the role of intra-host reproduction in the evolution of pathogen virulence, whereas the role of extra-host survival has received less attention. Here we examined the evolution of free-living virion survival in RNA virus populations differing in their histories of host-use. To do so, we used lineages of vesicular stomatitis virus (VSV) that were experimentally evolved in laboratory tissue culture for 100 generations on cancer-derived cells, non-cancerous cells, or alternating passages of the two host types. We observed that free-living survival improved when VSV populations specialized on human cancer-derived (HeLa) cells, whereas this trait was not associated with selection on non-cancer cells or combinations of the cell types. We attributed this finding to shorter-lived HeLa monolayers and/or rapid cell-to-cell spread of viruses on HeLa cells in tissue culture, both of which could select for enhanced virus stability in between host-cell replenishment. We also showed evidence that increases in virion survival were associated with decreases in virulence, suggesting a trade-off between survival and virulence for the VSV populations on one cell type. Our results shed new light on the causes and consequences of "sit and wait" infection strategies in RNA viruses.