Purpose: Gestational nutrition has an impact on the growth and development of the fetus.Choline (C) and docosahexaenoic acid (DHA) are important and essential nutrients for humans that play a role in the structural integrity of the membranes as well as signalling.C is used in the synthesis of phosphatidylcholine, and cell membranes are highly enriched with DHA. The dietary intake of C or DHA during pregnancy directly influences fetal development. Currently, there is no evidence to prove the effectiveness of the combined dietary supplementation of both C and DHA during gestation on developmental outcomes in the offspring. Methods: The current study was designed to assess the physical, sensory, and motor development of rat pups born to mothers supplemented with C and/or DHA during the entire gestational period. Pregnant rat dams were divided into the following five groups:Normal control (NC), Saline control (SC), Choline (C), DHA, and Choline+DHA (C+DHA).The NC dams did not receive any supplementation during the entire gestation period. The experimental groups were supplemented with Saline, C, and/or DHA, respectively, during the entire gestation (E0 to delivery). Results: Rat pups (n = 6/group) exposed to combined C and DHA showed significant improvement in birth weight, fur development, eye-opening as well as weight gain on the 7th, 14th, and 21st postnatal day and pinnae detachment (assessed from birth to postnatal day 21) when compared with age-matched NC, SC or C or DHA pups. Further, significant reflex responses were observed in visual placing and bar holding of pups exposed to both C and DHA, whereas the differences in surface righting, negative geotaxis, and grasping reflexes were not significant between the groups. Conclusion Gestational supplementation of both C and DHA rather than either of them alone is better in enhancing developmental outcomes in rat pups.