In vitro conservation of germplasm is considered the best alternative for preserving medicinal plants, enabling the multiplication and use of specific genotypes and contributing to the reduction of genetic erosion caused by anthropogenic or environmental impacts. These plants constitute an invaluable reservoir of substances with pharmacological activities and are, for this reason, overexploited, generally in an unsustainable manner, whether for popular, commercial, or research uses. Although the pharmaceutical industry produces many drugs, the chemical synthesis of complex molecules is not always possible. This chapter addresses the in vitro conservation of Petiveria alliacea, a tropical herbaceous species traditionally used as an analgesic, sedative, or abortifacient in folk medicine, as well as in religious rituals, owing to its neurological effects, indicating the great potential for the pharmacological use of bioactive substances synthesized in the plant against various human and animal pathologies, including neoplasms. This species has received much attention from researchers because of its biological activities, generating very complete knowledge about its phytochemical profile and the mechanisms of action of its main components, but conservation actions are scarce, being carried out, generally, at home in small gardens or in pots, by those who use or sell them. Research focused on in vitro conservation includes the establishment and standardization of micropropagation and cryopreservation protocols, as well as efficient systems for genetic monitoring of preserved material in addition to phytochemical studies and the biological effects of plants in vitro. Providing appropriate methodologies for the production and long-term conservation of the genetic resources of this species can contribute to the recovery of impacted natural populations, in addition to providing material for phytochemical and pharmacological research.