Purpose: To clarify the optimal dare of embryo transfer (ET), we retrospectively analyzed the relationship between the day of ET and the outcome in human in vitro fertilization and embryo transfer (IVF-ET). Method: Of a total of 307 human IVF-ET cycles performed at Kyoto University Hospital between January 1990 and March 1994, we focused on 207 cases of IVF-ET cycles in which two or three good-quality embryos were transferred. These 207 IVF-ET cycles consisted of 54 Day 2 ET cycles, 79 Day 3 ET cycles, 46 Day 4 ET cycles, and 28 Day 5 ET cycles. We compared the pregnancy and live-birth (plus ongoing pregnancy) rates among these four ET groups. Results: The pregnancy rates of ET on Days 2 to 4 were not significantly different, whereas Day 5 ET produced a significantly lower pregnancy rare (Day 2, 29.6%; Day 3, 32.9%; Day 4, 30.4%; Day 5, 10.7%). Similar results were obtained for the live-birth (plus ongoing pregnancy) rates (Day 2, 20.3%; Day 3, 18.9%; Day 4, 17.9%; Day 5, 7.1%). Conclusions: These results suggest that the day of ET does not fundamentally affect the pregnancy rate in human IVF-ET provided that transfer is made before Day 5.