A captive 9-yr-old female Siberian tiger (Panthera tigris altaica) at the Lisbon Zoo was treated with equine chorionic gonadotropin followed 80 hr later by human chorionic gonadotropin to stimulate follicular development and ovulation, respectively. Forty-one hours after the second hormone administration, the tiger was subjected to transvaginal artificial insemination with diluted fresh semen containing 500 × 10⁶ motile spermatozoa obtained from an 8-yr-old male Siberian tiger via electroejaculation. After anesthesia, the female tiger was placed in an inclined position with the hind limbs elevated during and after the insemination and was maintained in this position for 15 min to minimize semen reflux. An anesthetic reversal agent was then administered. After a 103-day gestation, the female gave birth to three apparently healthy cubs that survived 24–48 hr. These results demonstrate that transvaginal artificial insemination can be successfully performed in the Siberian tiger to produce full-term offspring.