Rodents have a well-developed visceral yolk sac (VYS) that acts as an active region for metabolic exchange and nutrition uptake until final fetal development in rats and may be affected by diabetes . Using Wistar rats, diabetes was induced by a single injection of alloxan at gestational day 8 (8 gd) and at 15 gd the collection of VYS was made. Flow cytometry was performed for VYS cell characterization, determination of mitochondrial activity, cell proliferation, DNA ploidy, cell cycle phases and caspase-3 activity. Fetal weight was reduced in the diabetic group. CD34, CCR2, and OCT3/4 expressions were significantly reduced, and CD90, CD117, and CD14 expressions were increased in the diabetic group. VYS cells with inactive mitochondria, activated caspase-3, and low proliferation were present in the diabetic condition. Severe hyperglycemia due to maternal diabetes was shown to have negative effects on pregnancy, VYS viability, and cell marker expression.