The production of new queens in honey bee colonies is one of the most important determinants of reproductive success, and it involves cooperative behavior among hundreds or thousands of workers. Colony members are generally expected to benefit by optimizing the reproductive traits of prospective replacement queens, but potential conflicts of interest among colony members could result in suboptimal queens. We studied the degree to which colonies regulate adult queen traits by controlling access to developing queens that survived from pupation to adulthood. We also searched for evidence of strong conflict among patrilines by comparing the contribution of patrilines to new queens and new workers, although we found no evidence for the existence of significantly queen-biased patrilines or for any association between patriline contribution to new queens and queen traits. However, adult queens emerging from cells accessible to workers were larger in terms of wet mass and thorax width compared to adult queens emerging from cells that were not accessible to workers. These results suggest that colonies regulate queen quality traits by curtailing low-quality queens from fully developing, which is further evidence that cooperation predominates over potential conflict within honey bee colonies.