To investigate risk factors of gestational diabetes mellitus (GDM) during assisted reproductive technology (ART) procedures. A total of 1022 patients were included in this retrospective cohort study from January 1, 2014 to August 31, 2017. While patients were divided into two groups: the non- GDM group and the GDM group. There was no significant difference in basal FSH, AFC, infertility years, gestational age, number of fetus, method of fertilization, and reason of infertility between the two groups. However, age, BMI, and fresh cycle were verified to be association with GDM by using logistic regression model. During the process of controlled ovarian hyperstimulation (COH), estradiol (E-2) was significantly lower in the GDM group. The incidence of GDM was highest when E-2 level less than 200 pg/mL of per oocyte. Our study showed maternal fundamental factors had greater impacts on subsequent GDM.