To investigate whether children who consumed infant formula supplemented with long-chain polyunsaturated fatty acids (LCPUFAs) had a more favourable cardiovascular profile than children who consumed formula without these fatty acids, we used the Wheezing Illnesses Study Leidsche Rijn, a birth cohort that included 2,468 newborns between 2001 and 2014. Data on infant feeding were obtained by questionnaires. At age 5, blood pressure, carotid intima-media thickness (CIMT), and carotid distension were measured. We used multivariable linear regression analysis to compare levels of cardiovascular markers in formula-fed children born before and after the LCPUFA supplementation. To account for secular trends, we compared levels of cardiovascular markers in a control group of breastfed children from the same cohort born before and after the supplementation. Formula-fed children born after the LCPUFA supplementation (n=48) had no different systolic blood pressure (-2.58mmHg, 95% confidence interval, CI [-5.5, 0.30]), diastolic blood pressure (-0.13mmHg, 95% CI [-2.3, 2.1]), or carotid distension (24.8MPa-1 , 95% CI [-47.1, 96.6]) and had a higher CIMT (18.6μm, 95% CI [3.7, 33.5]) than formula-fed children born before the supplementation (n=163). In the control group, children born after the LCPUFA supplementation (n=98) had no different systolic- or diastolic-blood pressure, or CIMT, and a higher carotid distension than children born before the supplementation (n=142). In conclusion, children who consumed infant formula supplemented with LCPUFAs did not have a more favourable cardiovascular profile in early childhood than children who consumed formula without LCPUFAs.