Ye Htut died from health problems related to old age at a hospital in the main city of Yangon on Wednesday, family members said.
The Thirty Comrades were led by Gen. Aung San, father of democracy leader Aung San Suu Kyi. During World War II, the men went to Japan for training to fight British colonizers in what was then known as Burma. Aung San later negotiated independence from Britain, but was assassinated before that occurred in 1948.
Ye Htut, who served in the Myanmar army until independence, went underground soon afterward, joining the armed struggle of the banned Burma Communist Party.
He laid down his weapons in 1963 to join the ruling party of then-dictator Gen. Ne Win, but was purged several years later in an inner-party struggle, according to his eldest son, Kyaw Kyaw.
Ye Htut was involved in the 1988 pro-democracy movement.
Tin Oo, a former chief of staff and veteran of Aung San Suu Kyi's National League for Democracy party, expressed his condolences, saying he had great respect for those who helped the country fight for independence.
"I am very sad to hear about the death of the last surviving member of the Thirty Comrades," he said.
Ye Htut "served as a patron of the Patriotic Old Comrades league—a group formed by retired army leaders during the peak of the 1988 uprising.
Ye Htut is survived by his two sons.