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Methods: In this retrospective cohort study, 203 out of 1686 women with twin pregnancies received natural progesterone (200 mg/day between gestational weeks 16 + 0 and 36 + 0) vaginally for >= 4 weeks. The control group consisted of 1483 women with twin pregnancies without progesterone administration. Pearson's Chi squared test, Fisher's exact test, and Student'st-test was used to compare differences between the control and the progesterone-treated groups. A multivariate binary logistic regression was performed to assess relative independent associations on the dependent outcome of GDM incidence. Results: Vaginal progesterone treatment in twin pregnancies had no significant influence on developing GDM (p= 0.662). Higher pre-pregnancy BMI (OR 1.1;p< 0.001), GDM in previous pregnancy (OR 6.0;p< 0.001), and smoking during pregnancy (OR 1.6;p= 0.014) posed an increased risk for developing GDM. 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