Spatial release from masking (SRM) was tested within the first week of fitting and after 12 weeks hearing aid use for unilateral and bilateral adult hearing aid users. A control group of experienced hearing aid users completed testing over a similar time frame. The main research aims were (1) to examine auditory acclimatization effects on SRM performance for unilateral and bilateral hearing aid users, (2) to examine whether hearing aid use, level of hearing loss, age or cognitive ability mediate acclimatization, and (3) to compare and contrast the outcome of unilateral versus bilateral aiding on SRM. Hearing aid users were tested with and without hearing aids, with SRM calculated as the 50% speech recognition threshold advantage when maskers and target are spatially separated at ±90° azimuth to the listener compared to a co-located condition. The conclusions were (1) on average there was no improvement over time in familiar aided listening conditions, (2) there was large test-retest variability which may overshadow small average acclimatization effects; greater improvement was associated with better cognitive ability and younger age, but not associated with hearing aid use, and (3) overall, bilateral aids facilitated better SRM performance than unilateral aids.