Background and Objectives: The skin barrier protects skin against a harmful environment. Its function includes an antimicrobial barrier as well as a physical barrier. Stress is harmful to the skin barrier and there are many studies on this, but there are few about the effect of stress during pregnancy on the skin barrier of offspring. The aim of this study was to investigate the effect of stress during pregnancy on the skin barrier of offspring by examination with the naked eye, cortisol, weight, TEWL and histologic tests. Materials and Methods: Male hairless mice ten weeks old were coupled with females for three days. After twelve days the females were divided into two groups. We stressed one group and not the other group. The offspring from the non-stressed (N group) and stressed (S group) grew up until four weeks. The S group was exposed to chronic mild stress using the chronic stress model protocol modified method of Wilner and Towell for a week. We made examinations with the naked eye, assessment of cortisol, weight, TEWL, and histologic test to contrast the S group with the N group. Statistical analysis was performed by using Mann-Whitney and Friedman test. Statistical significance was achieved if the probability was less than 5% (p<0.05). Results: 1. By making an examination with the naked eye, S group showed papules, creases and dryness on their skin, but N showed no change compared with normal skin. 2. There was no statistically significant difference between S group and N group in cortisol (p>0.05). 3. There was no statistically significant difference between S group and N group in weight. N group had less weight than S group (p<0.05). 4. There was no statistically significant difference between S group and N group in TEWL (p>0.05). 5. There was no statistically significant difference between S group and N group in TEWL recovery (p>0.05). 6. In TEWL and TEWL recovery, there was statistically significant difference by time between S group and N group (p<0.005). Histologically, S group showed a thicker epidermis and epidermal crest than N group. Conclusions: We found that stress during pregnancy has negative effects on the skin barrier of offspring. Though there were no statistically significant differences between S group and N group in TEWL recovery, we found evidence that the recovery of skin barrier function is interrupted by stress during pregnancy by examination with both the naked eye and histology.