Tu Youyou was born in Ningbo, Zhejiang province, China, a city known for its rich history and intellectual tradition. Her name, "Youyou", was inspired by a line from the Book of Odes, a classic collection of Chinese poetry, reflecting her parents' hope for a peaceful and harmonious life for her.

Tu Youyou enrolled at Peking University's School of Pharmaceutical Sciences, where she majored in pharmaceutical chemistry. This period of formal education provided her with a foundational understanding of drug development, extraction techniques, and the principles of chemistry necessary for her future work.