It is known that more than 180 species of mushrooms contain psilocybin or its derivative psilocin, and the fungi have a long history of use in Mesoamerican spiritual and religious rituals. Imagery found on prehistoric murals and rock paintings of modern-day Spain and Algeria suggests that human usage of psilocybin mushrooms predates recorded history. In Mesoamerica, the mushrooms had long been consumed in spiritual and divinatory ceremonies before Spanish chroniclers first documented their use in the sixteenth century.

There are extensive accounts of pre-Columbian psilocybin use among the Mayan and Aztec cultures of Mesoamerica, namely in Mexico and Guatemala. After conquering these areas in the 15th and 16th centuries, the Spanish forbade psychedelic mushroom use by indigenous peoples, regarding it as a savage and uncivilized cultural practice. Despite this, the indigenous shamans ignored Spanish law in secret for over 400 years and continued to use and administer these mushrooms to preserve their cultural heritage.

The Mazatec Shamans are known for their ritual use of psilocybin mushrooms, psychoactive morning glory seeds, and Salvia divinorum. In their rituals, Mazatec shamans use fresh Salvia divinorum leaves. Ritual use traditionally involves being in a quiet place after ingestion of the leaf—the Maztec shamans say that "La Maria (S. divinorum) speaks with a quiet voice."

In 1959, the Swiss chemist Albert Hofmann isolated the active principle psilocybin from the mushroom Psilocybe mexicana. Hofmann's employer Sandoz marketed and sold pure psilocybin to physicians and clinicians worldwide for use in psychedelic psychotherapy. Although the increasingly restrictive drug laws of the late 1960s curbed scientific research into the effects of psilocybin and other hallucinogens, its popularity as an entheogen (spirituality-enhancing agent) grew in the next decade, owing largely to the increased availability of information on how to cultivate psilocybin mushrooms.