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Background: Professional quality of life pertains to the balance between compassion fatigue and compassion satisfaction. In recent years, there was an increase in compassion fatigue among medical staff due to the pandemic, all over the world, while compassion satisfaction was reported at a moderate level. Objectives: The aim of this study was to identify the relationship between workplace humour and professional quality of life among medical staff, as well as the moderating role of compassion in this relationship. Method: The sample consisted of 189 participants (Mage = 41.01; SD = 9.58). Of the total sample, 57.1% are physicians, 32.3% are nurses and 6.9% are clinical psychologists. The participants completed scales measuring compassion, workplace humour, and professional quality of life. Results: The results showed that self-enhancing and affiliative humour were positively related, while self-defeating humour was negatively related to compassion satisfaction. Burnout and secondary traumatic stress were negatively related to self-enhancing humour and positively related to self-defeating humour. Compassion moderated the relationship between affiliative humour and secondary traumatic stress. Conclusions: Encouraging coping strategies based on adaptive humour (i.e. affiliative humour, self-enhancing) and raising awareness about negative humour strategies (i.e. self-defeating) could contribute to an increase of quality of life among healthcare providers. Another conclusion derived from the present study sustains that compassion is a valuable personal resource positively related to compassion satisfaction. Compassion also facilitates the relationship between affiliative humour and low secondary traumatic stress. Thus, encouraging compassionate skills could be beneficial for the optimal professional quality of life.