AbstractMaternal nutrition is pivotal for offspring’s growth and development. Insufficient or unbalanced nutrition may cause osteoporosis and other diseases. Protein and calcium are essential dietary nutrients for offspring’s growth. However, the optimal contents of protein and calcium in maternal diet remain unclear. In the present study, we set four different protein and calcium content-pregnancy nutrition groups, including normal full-nutrient (Normal), low protein and low calcium (Pro−; Ca−), high protein and low calcium (Pro+; Ca−), and high protein and high calcium groups (Pro+; Ca+), to evaluate the weight gain of maternal mice as well as the weight, bone metabolism, and bone mineral density of offspring mice. When the vaginal plug is found, the female mouse will be kept in a single cage and fed with corresponding feed until delivery. The findings demonstrate that Pro−; Ca− diet affects the growth and development of offspring mice after birth. In addition, a low-calcium diet inhibits the growth of embryonic mice. Collectively, the present work further confirms the importance of protein and calcium in the maternal diet and deeply suggests their respective roles in different development stages.