The last 13 animals in what had once been a group of 54 will be moved to U.S. zoos or sanctuaries from their current home in the hills above campus.
Researchers have become "extremely attached" to the animals, which are often stereotyped as obnoxious scavengers, said George Bentley, integrative biology professor and director of the field station housing the animals.
"They're comical, charismatic, social, and very intelligent animals," he said. "They're fantastic to watch and incredibly intriguing."
Hyenas in the colony were used as models for characters in the Disney movie "The Lion King."
The colony grew from cubs first brought from Kenya's Maasai Mara region in 1985. Scientists have studied their social structure and aggression, striking vocalizations and the females' unusual anatomy.
The clitoris of the female hyena resembles male genitalia, leading many scientists to theorize that the animals were hermaphrodites. This was disproved by integrative biology professor Stephen Glickman, who had helped bottle-feed the first batch of Berkeley's hyena cubs.
But Glickman did find that the females give birth, urinate, and mate through their "pseudo penis," and dominate males.
Justin Goodman, director of laboratory investigation for the People for the Ethical Treatment of Animals, hailed the decision to move the hyenas.
"Any time lab animals are released is a time to celebrate," Goodman said.