Humphrey was born in Oak Park, Illinois, but grew up in Chicago, Illinois. She was the daughter of Horace Buckingham Humphrey, a Journalist and one-time hotel manager, and Julia Ellen Wells, who had trained as a concert Pianist. She was a descendant of pilgrim william Brewster who arrived on the Mayflower in 1620. In Chicago, with the encouragement of her mother, she studied with eminent ballet masters as well as with Mary Wood Hinman, who taught dance at her school, the Francis Parker School. While still at high school she undertook a concert tour of the western states as a Dancer, with her mother as accompanist, in a group sponsored by the Santa Fe Railroad for its Workman’s Clubs.

Partly due to financial concerns Humphrey opened her own dance school, with her marg as manager and Pianist, in 1913 at the age of 18. It was a great success, offering classic, gymnastic and ballroom dance for children and ballroom dance for young adults.

In 1917, at the instigation of Mary Wood Hinman, she moved to California and entered the Denishawn School of Dancing and Related Arts where she studied, performed, taught classes and learned choreography. Her creations from this era, Valse Caprice (Scarf Dance), Soaring, and Scherzo Waltz (Hoop Dance) are all still performed today. She remained involved for the next decade. Humphrey toured the Orient for two years followed by a successful career in American vaudeville theaters.

Humphrey's most prominent works include Color Harmony (1928), Water Study (1928), Drama of Motion (1930), and The Shakers (1931). Color Harmony (1928) was her first independent concert after leaving Denishawn in 1928. She and Charles Weidman presented the work with "Weidman as a slivery figure representing the artistic intelligence that organizes the mingled colors of the spectrum into a harmonious design." Water Study (1928) was Humphrey's experiment of dancing without music. She wanted the Dancers to move to their natural breathing patterns as they represented the natural movement of water. Drama of Motion (1930) was the next step in her experiment of dancing without music. She wanted dance to be an art that could stand on its own without the need of music or emotion and concentrated on the formal elements of movement such as design, rhythm, and dynamics. Her best known work however, brought music and emotion back into play. The Shakers (1931) was inspired by a Christian sect known as Shakers. They required members to be celibate and are known for shaking during times of worship. To make sure their members remain celibate they segregate the sexes during rituals. In Humphrey's choreography she incorporates shaking movements to represent their sexual repression as well as the idea of being shaken clean of sin.

Humphrey was a participant of the Federal Dance Project (FDP), created in the 1930s as part of Franklin Delano Roosevelt's Second New Deal. FDP was the first national program created to financially support dance and Dancers.

Short in stature, Doris was no taller than 5'3" and had a slender build. In 1932 she married Charles Woodford (a merchant.) She had one child, a son named Charles Humphrey Woodford (born 1934.)

Humphrey expanded her choreographic work to Broadway in 1933 with School for Husbands and again the following year with Life Begins at 8:40.

During the 1940s, Humphrey spent significant time with José Limón, one of her former students. After she retired from performing in 1944, due to arthritis, she became artistic Director for the José Limón Dance Company and created a number of works for the company, including Day on Earth, Night Spell, Ruins and Visions, and Lament for Ignacio Sanchez Mejias. Humphrey’s dance style was carried on and developed further by Limón and his dance company.

In 1952 Humphrey started directing a new dance company for children called The Merry-Go-Rounders.

Shortly after her death in 1958, Humphrey's book, The Art of Making Dances, in which she shared her observations and theories on dance and composition, was published. In the introduction she observed that ballet had changed radically in the 20th century. "Suddenly the dance," she said, "the Sleeping Beauty, so long reclining in her dainty bed, had risen up with a devouring Desire.” She believed in emotions and movement moving “from the inside out”, but she also believed in working abstractly where specific events and characters were not illustrated in a way that made sense. For Example, “she believed that the concept of democracy was more convincingly conveyed by a fugue uniting four different themes than by a woman in red, white and blue”.

Humphrey was inducted into the National Museum of Dance's Mr. & Mrs. Cornelius Vanderbilt Whitney Hall of Fame in 1987.

The book, Days on Earth: The Dance of Doris Humphrey, by Marcia B. Siegel was published in 1993. Siegel makes a case for Humphrey being one of the more important figures of modern dance.

One of her last pieces, Dawn in New York, showed the strengths Limón demonstrated throughout her career – her mastery of the intricacies of large groups and her emphasis on sculptural shapes. Humphrey was on the original faculties of both The Bennington School of the Dance (1934) and The Juilliard School (1951), both directed by Martha Hill.

Her choreography from these early years includes Air for the G String, Water Study, Life of the Bee, Two Ecstatic Themes, and The Shakers. Unlike the Denishawn approach in choreography, finding inspiration from abroad, Humphrey sought inspiration from within her home, America. The Shakers, about the 18th century American religious group, is a notable Example of finding inspiration from America.

Humphrey’s theory explored the nuances of the human body's responses to gravity, embodied in her principle of "fall and recovery". She called this “the arc between two deaths”. At one extreme, an individual surrenders to the nature of gravity; at the other, one attempts to achieve balance. Through the fall and recovery principle, Humphrey is able to illustrate emotional and physical climax of struggling for stability and submitting to the laws of gravity.