Climate-induced fire regime shifts may reduce post-fire resilience of black spruce-dominated (BS; Picea mariana) North American boreal forests. While post-fire vulnerability of immature BS stands has been extensively studied, no study has evaluated simultaneous effects of fire severity and seasonality on the post-fire regeneration of mature (> 60-year-old) BS stands. This study aims to quantify post-fire regeneration levels of BS and co-occurring tree species to assess ecosystem recovery and possible loss of resilience due to regeneration failure. We analyzed effects of seed bank conditions, fire regime characteristics (fire severity and seasonality), and seedbed conditions on BS post-fire regeneration in mature forests in Quebec, Canada. Post-fire regeneration density was extensively surveyed across ~50 400 km2 through a network of 536 plots that were distributed in 21 fires, which burned between 1995 and 2016. One-third of plots failed to regenerate (< 1750 conifer seedlings/ha) at levels adequate to produce closed-crown forest, whereas one-fifth experienced compositional changes, mainly towards jack pine (JP; Pinus banksiana) dominance. Pre-fire basal area of BS and living Sphagnum ground cover increased BS post-fire regeneration, whereas high-severity crown fires and spring fires reduced it. These findings suggest that mature BS-dominated forests may lose resilience in response to high-severity and spring fires. Given the projected increase in fire severity, and the extension towards an early-fire season in response to climate change, our study suggests that post-fire regeneration failure may become more frequent over the coming decades, with potential negative consequences on ecosystem services that are provided by BS-dominated boreal forests.