The proper nutritional status during the hormone-sensitive growth phases preceding first parturition can affect mammary development and subsequent lactation performance. We developed a compensatory nutrition regimen (CNR), which is designed to stimulate mammary growth by exploiting the biological characteristics of the energy restriction and compensatory growth phenomenon. In the present study, we examined the effect of compensatory growth induced only once during late gestation upon mammary development and subsequent lactation potential over 2 lactation cycles. Female rats were mated and randomly assigned to either the control or the CNR group. Control rats were offered the control diet (AIN-93G) throughout the experiment. CNR rats were subjected to 40% energy restriction during the first 10 d of gestation followed by free access to the control diet for the remainder of the experiment. Dams on the CNR produced 14% more milk than control dams (P = 0.12). Mammary cell proliferation rates were approximately 46% (P < 0.05) and 27% (P = 0.07) higher in the CNR group than in the control during late gestation and early lactation of the first lactation cycle, respectively. Caspase-3 enzyme activity was decreased 15% (P < 0.05) and 22% (P = 0.11) in mammary tissues from the CNR group compared with that from the controls during the first and second lactation cycles, respectively. These results indicate that compensatory growth induced only once during late gestation increases mammary cell proliferation and differentiation and decreases regression of mammary cells throughout consecutive lactation cycles.