Background:Milk formulas for artificially reared (AR) rat pups are mostly based on complex cow's milk products, prepared by laborious and time-consuming processes. The aim of this study was to develop a simplified procedure for preparing a refined formula and to examine its influences on gut and brain development.Methods:The formula comprised a combination of purified cow's casein and whey proteins, five kinds of edible oil, minerals, and vitamins. Detailed analyses showed that the composition of macro- and micro-nutrients, osmolarity, and pH of the new formula closely resembled those of rat's milk. Rat pups, each with an intragastric cannula implanted at age 5 days, were artificially reared for the following 10-15 days.Results:The body weight gain of AR pups matched that of mother-reared (MR) pups. Histoplanimetrical analyses showed that the small intestine in AR pups was more developed in relation to area of a transverse section, number and length of villi, and thickness of tunica muscularis than that of MR pups. Fat components in the formula influenced the fatty acid composition and the cholesterol-to-phospholipid ratio in the small intestinal microvillus membrane (MVM) of AR pups, but not the MVM fluidity. Brain weight was not significantly different between the two groups at age 15-20 days.Conclusion:This formula is useful for artificial rearing of rats and for identifying dietary components contributing to metabolic adaptation during the suckling period.