This study considers the effect of crossbreeding and gender (barrows; gilts) on meat quality and intramuscular and subcutaneous fatty acid composition in pork. The sire lines included Large White (LW), Duroc (D) and Pietrain (P) and the dam line was Landrace (LR)×LW, producing LW×(LR×LW), D×(LR×LW) and P×(LR×LW). Pork samples were removed from Longissimus dorsi (LD) and Semimembranosus (SM) muscles and subcutaneous fat (SCF). There were no important differences in meat quality parameters but D×(LR×LW) had the highest percentage of intramuscular fat. Castrated males had more intramuscular fat and more intense meat colour than female pigs. The Large White and Duroc sire line had saturated fatty acids (SFA) in SM, whereas the Pietrain sire line was significantly higher than Duroc sire line in the concentration of polyunsaturated fatty acids (PUFA), PUFA/SFA (P/S) and the n-6/n-3 ratio. The concentrations of SFA and monounsaturated fatty acids (MUFA) in SCF were significantly higher in LW×(LR×LW) and P×(LR×LW), respectively. No differences were found in the percentage of PUFA, P/S and n-6/n-3 ratio between D×(LR×LW) and P×(LR×LW). Female pigs had the most polyunsaturated intramuscular and subcutaneous fat. The results demonstrate small differences in fatty acid compositions among sires.