Beginning in 1916, Tarsila studied painting in São Paulo. Later she studied drawing and painting with the academic Painter Pedro Alexandrino. These were all respected but conservative teachers. Because Brazil lacked a public art museum or significant commercial gallery until after World War II, the Brazilian art world was aesthetically conservative and exposure to international trends was limited. Tarsila studied in Paris from 1920 through June of 1922 at the Académie Julian with Emile Renard.

Returning to São Paulo in 1922, Tarsila was exposed to many things after meeting Anita Malfatti, Oswald de Andrade, Mário de Andrade, and Menotti Del Picchia. Prior to her arrival in São Paulo from Europe, the group had organized the Semana de Arte Moderna ("Week of Modern Art") during the week of February 11–18, 1922. The event was pivotal in the development of modernism in Brazil. The participants were interested in changing the conservative artistic establishment in Brazil by encouraging a distinctive mode of modern art. Tarsila was asked to join the movement and together they became the Grupo dos Cinco, which sought to promote Brazilian culture, the use of styles that were not specifically European, and the inclusion of things that were indigenous to Brazil.

Oswald de Andrade, who had become her traveling companion, accompanied her throughout Europe. Upon returning to Brazil at the end of 1923, Tarsila and Andrade then traveled throughout Brazil to explore the variety of indigenous culture, and to find inspiration for their nationalistic art. During this period, Tarsila made drawings of the various places they visited which became the basis for many of her upcoming paintings. She also illustrated the poetry that Andrade wrote during their travels, including his pivotal book of poems entitled Pau Brasil, published in 1924. In the manifesto of the same name, Andrade emphasized that Brazilian culture was a product of importing European culture and called artists to create works that were uniquely Brazilian in order to "export" Brazilian culture, much like the wood of the Brazil tree had become an important export to the rest of the world. In addition, he challenged artists to use a modernist approach in their art, a goal they had strived for during the Semana de Arte Moderna in São Paulo.

During this time, Tarsila's colors became more vibrant. In fact, she wrote that she had found the "colors I had adored as a child. I was later taught they were ugly and unsophisticated." Her initial painting from this period was E.C.F.B.(Estrada de Ferro Central do Brasil), (1924). Furthermore, at the time, she had an interest in industrialization and its impact on society.

In 1926, Tarsila married Andrade and they continued to travel throughout Europe and Middle East. In Paris, in 1926, she had her first solo exhibition at the Galerie Percier. The paintings shown at the exhibition included São Paulo (1924), A Negra (1923), Lagoa Santa (1925), and Morro de Favela (1924). Her works were praised and called "exotic", "original", "naïve", and "cerebral", and they commented on her use of bright colors and tropical images.

Tarsila's first painting during this period was Abaporu (1928), which had been given as an untitled painting to Andrade for his birthday. The subject is a large stylized human figure with enormous feet sitting on the ground next to a cactus with a lemon-slice sun in the background. Andrade selected the eventual title, Abaporu, which is an Indian term for "man eats", in collaboration with the poet Raul Bopp. This was related to the then current ideas regarding the melding of European style and influences. Soon after, Andrade wrote his Anthropophagite Manifesto, which literally called Brazilians to devour European styles, ridding themselves of all direct influences, and to create their own style and culture. Instead of being devoured by Europe, they would devour Europe themselves. Andrade used Abaporu for the cover of the manifesto as a representation of his ideals. The following year the manifesto's influence continued, Tarsila painted Antropofagia (1929), which featured the Abaporu figure together with the female figure from A Negra from 1923, as well as the Brazilian Banana leaf, cactus, and again the lemon-slice sun.

In 1929, Tarsila had her first solo exhibition in Brazil at the Palace Hotel in Rio de Janeiro, and was followed by another at the Salon Gloria in São Paulo. In 1930, she was featured in exhibitions in New York and Paris. Unfortunately, 1930 also saw the end of Tarsila and Andrade's marriage. This brought an end to their collaboration.

The remainder of her career she focused on social themes. Representative of this period is the painting Segundo Class (1931), which has impoverished Russian men, women and children as the subject matter. She also began writing a weekly arts and culture column for the Diario de São Paulo, which continued until 1952.

In 1938, Tarsila finally settled permanently in São Paulo where she spent the remainder of her career painting Brazilian people and landscapes. In 1950, she had an exhibition at Museum of Modern Art, São Paulo where a reviewer called her "the most Brazilian of Painters here, who represents the sun, birds, and youthful spirits of our developing country, as simple as the elements of our land and nature…. She died in São Paulo 1973. Tarsila's life is a mark of the warm Brazilian character and an expression of it tropical exuberance."

In 2018 MoMA opened a solo exhibition of her work, the eighth retrospective on Latin America artists following exhibitions on Diego Rivera, Cândido Portinari, Roberto Matta, Manuel Álvarez Bravo, Armando Reverón, José Clemente Orozco and Joaquín Torres García.