Three-dimensional printed denture base resins are relatively new materials, and their properties need to be thoroughly investigated to assess whether they can be used clinically. The aim of the current study was to evaluate the colour stability of 3D printed and conventional denture base resins after immersion in different staining solutions. A total of 200 specimens were manufactured from two types of materials: 3D printed dental resin NextDent Denture 3D+ (NextDent, 3D Systems, the Netherlands) and heat-polymerized PMMA Vertex (3D Systems, the Netherlands), which were immersed in four types of colourants—artificial saliva, coffee, red wine and coke (n = 25). For measuring the colour changes (CIE-L*a*b* system), a SpectroShade Micro spectrophotometer (SpectroShade, Oxnard, CA, USA) was used. After seven days (T1), 14 days (T2) and 21 days (T3), the mean ∆E values were calculated and compared by the Bonferonni post hoc test. The data were processed using the statistical software SPSS 26. The level of significance for rejecting the null hypothesis was fixed at p < 0.05. The highest mean values for ∆E were found for both types of dental resin in red wine, and the lowest mean values for ∆E were found for 3D printed specimens in artificial saliva. The 3D printed denture base resin demonstrated better colour stability than the conventional acrylic materials. The staining effect correlated with the immersion time, with the red wine and coke having the strongest chromogenic impact and the period with the highest colour changes being 21 days.