Diapause control and colony initiation are among the major problems encountered in the rearing of bumble bee colonies in small-scale rearing. In this study, we used Bombus terrestris queens obtained from commercial colonies to investigate (1) the diapause survival in virgin and mated queens, (2) the diapause and colony initiation performance of mated bumble bee queens in relation to the pre-diapause weight and (3) the effect of pupae’s sex on colony initiation. We found that diapause survival is negatively affected by mating and by the low pre-diapause weight, but first egg deposition and development of the first adult worker were delayed in heavy queens. We found no significant differences in the egg-laying success in relation to pupae’s sex; however, queens stimulated with queen pupae laid more eggs per cell and developed a first brood larger than those stimulated with male pupae. Our results can be useful in small-scale rearing, including the rearing of wild queens for conservation purposes.