Background: Advanced glycation end products (AGEs) are tissue proteins that accumulate with age and in diabetes mellitus (DM). AGEs can be measured by the AGE-Reader (DiagnOptics Technologies BV, Groningen, The Netherlands), which measures skin autofluorescence (SAF). SAF has been suggested as a measure to screen for undiagnosed DM or impaired glucose tolerance. SAF has never been investigated in gestational DM (GDM). Therefore we compared SAF at diagnosis in GDM patients with normal pregnancy. If SAF is elevated in GDM, future research could focus on the possible use of the AGE-Reader as a screening method for GDM. Methods: In this monocenter observational study SAF was measured in 60GDM patients at diagnosis and 44 pregnant women without diabetes. Results: SAF did not differ between GDM at diagnosis (mean [SD], 1.74 [0.31] arbitrary units) and normal pregnancy (1.76 [0.32] arbitrary units). SAF was lower in white European patients than in patients with other ethnicity. Conclusions: This first study of tissue AGE accumulation in pregnancy shows no differences in SAF between women with GDM at diagnosis and normal pregnancy. This is most likely due to mild severity and short duration of hyperglycemia in GDM at diagnosis, but it does not exclude potential differences in SAF later in pregnancy. However, the fact that no differences are detected at diagnosis makes it unlikely that the AGE-Reader can be developed as a screening method for GDM in the future. Furthermore, we found that ethnicity should be taken into account when measuring SAF.