The role of psychosocial and physical factors in the development of musculoskeletal pain has now been clearly shown. But it is not clear whether these factors contribute to specific regional musculoskeletal pain or to multisite pain. The main goal of this study was to assess the impact of work-related factors according to gender to the development of regional and multisite musculoskeletal pain. In all, 12,591 subjects (65% men) born in 1938, 1943, 1948 and 1953 participating in a French longitudinal prospective epidemiological survey in 1990-1995 (ESTEV) were eligible. Personal factors and work exposure were assessed by self-administered questionnaires. Statistical associations between chronic musculoskeletal pain (regional body site or multisite), personal and occupational factors were analysed using logistic regression modelling. The incidence of regional musculoskeletal pain and multisite pain in 1995 were respectively 17% and 25.6%. For women, highly repetitive movements predicted neck/shoulder pain, posture and vibrations predicted arm and low back pain, effort with tools predicted arm pain. For men, forceful effort and vibrations predicted neck/shoulder pain, posture and forceful effort predicted lower limb and low back pain, forceful effort and effort with tools predicted arm pain. Physical constraints (forceful effort or vibrations) were associated with multisite pain in both genders. Only for women, psychological elements are predictive risk factors of upper limb pain and of three or four painful anatomical sites. These results support the hypothesis that some physical and psychological work-related factors are predictive of regional or multisite musculoskeletal pain but differ between genders. Gender differences and risk factors of work-related musculoskeletal pain should be also taken into consideration to more effectively focus preventive measures.