Women treated with hemodialysis report lower health-related quality of life (HRQoL) compared with men. Whether this is related to sex-specific (biological) (e.g. under-dialysis due to body composition differences) or gender-specific (sociocultural) factors (e.g. greater domestic/caregiver responsibilities for women) is unknown. We examined the association between sex assigned at birth, gender score and HRQoL in individuals initiating conventional and incremental hemodialysis. In this prospective multi-center cohort study, incident adult hemodialysis patients were recruited between 1 June 2020 and 30 April 2022 in Alberta, Canada. Sex assigned at birth and gender identity were self-reported. Gender-related characteristics were assessed by self-administered questionnaire to derive a composite measure of gender. The primary outcome was change in Kidney Disease Quality of Life 36 physical (PCS) and mental (MCS) component scores after 3 months of hemodialysis. Sixty participants were enrolled (conventional hemodialysis: 14 female, 19 male; incremental hemodialysis: 12 female, 15 male). PCS improved from baseline with conventional (P=.01) but not incremental (P=.52) hemodialysis in female participants. No difference in MCS was observed by hemodialysis type in female participants. Gender score was not associated with changes in PCS in female participants, irrespective of hemodialysis type. Higher gender score was associated with increased MCS with incremental (P=.04), but not conventional (P=.14), hemodialysis (P=.03 conventional vs incremental) in female participants. No change in PCS or MCS was seen in male participants, irrespective of hemodialysis type or gender score. In this exploratory study, conventional hemodialysis was associated with improved PCS in female participants, while incremental hemodialysis was associated with improved MCS in female participants with more roles and responsibilities traditionally ascribed to women. Large prospective studies are required to further investigate these relationships.