Marek’s disease virus (MDV) is an important poultry pathogen which is controlled through widespread vaccination with avirulent and attenuated strains, but continued evolution of field viruses to higher virulence has required ongoing improvement of available vaccine strains, and these vaccine strains also offer an attractive platform for designing recombinant vector vaccines with cross-protection against MDV and additional pathogens. Recent reports of failures in vaccine licensing trials of positive controls to reach appropriately high levels of MD incidence prompted us to evaluate possible combinations of outbred specific pathogen-free (SPF) layer lines and alternative virulent challenge strains which could provide more consistent models for serotype-3 vectored vaccine development. Choice of layer line and virulent MDV challenge strain each contributed to the ability of a challenge model to reach 80 percent virulence in unvaccinated positive control groups in the majority of trials without overwhelming serotype-3 vectored vaccine protection in vaccinated groups. Conversely, reducing challenge virus dose by a factor of four, or vaccine dose by half, had no consistent effect across these models. Although MDV strain 617A had the most potential as an alternative to strains that are currently approved for licensing trials, no combination of layer line and challenge virus consistently met the goals for a successful challenge model in all study replicates, indicating that high variability is an inherent difficulty in MDV challenge studies, at least when outbred birds are used.