Oilseed supplementation is a strategy to improve milk production and milk composition in dairy cows; however, the response to this approach is inconsistent. Thus, the aim of this study was to evaluate the effect of oilseed supplementation on milk production and milk composition in dairy cows via a meta-analysis and meta-regression. A comprehensive and structured search was performed using the following electronic databases: Google Scholar, Primo-UAEH and PubMed. The response variables were: milk yield (MY), atherogenic index (AI), Σ omega-3 PUFA, Σ omega-6 PUFA, fat, protein, lactose, linoleic acid (LA), linolenic acid (LNA), oleic acid (OA), vaccenic acid (VA), conjugated linoleic acid (CLA), unsaturated fatty acid (UFA) and saturated fatty acid (SFA) contents. The explanatory variables were breed, lactation stage (first, second, and third), oilseed type (linseed, soybean, rapeseed, cottonseed, and sunflower), way (whole, extruded, ground, and roasted), dietary inclusion level, difference of the LA, LNA, OA, forage and NDF of supplemented and control rations, washout period and experimental design. A meta-analysis was performed with the “meta” package of the statistical program R. A meta-regression analysis was applied to explore the sources of heretogeneity. The inclusion of oilseeds in dairy cow rations had a positive effect on CLA (+0.27 g 100 g−1 fatty acids (FA); p < 0.0001), VA (+1.03 g 100 g−1 FA; p < 0.0001), OA (+3.44 g 100 g−1 FA; p < 0.0001), LNA (+0.28 g 100 g−1 FA; p < 0.0001) and UFA (+8.32 g 100 g−1 FA; p < 0.0001), and negative effects on AI (−1.01; p < 0.0001), SFA (−6.51; p < 0.0001), fat milk (−0.11%; p < 0.001) and protein milk (−0.04%; p < 0.007). Fat content was affected by animal breed, lactation stage, type and processing of oilseed and dietary NDF and LA contents. CLA, LA, OA and UFA, desirable FA milk components, were affected by type, processing, and the intake of oilseed; additionally, the concentrations of CLA and VA are affected by washout and design. Oilseed supplementation in dairy cow rations has a positive effect on desirable milk components for human consumption. However, animal response to oilseed supplementation depends on explanatory variables related to experimental design, animal characteristics and the type of oilseed.