The choice of explant and its preparation are most likely one of the most important biotic factors that determine the success of a tissue culture protocol when the explant is derived from ex vitro organs. This review assesses the disinfection protocols available for saffron (Crocus sativus L.; Iridaceae) explants in an attempt to ascertain the most suitable set of parameters that could ensure successful tissue culture in subsequent treatments. From a methodological perspective, two explant types are most commonly adopted in saffron biotechnology, namely the use of fresh field-grown shoots, or dormant corms. The latter is more recommended to minimize contamination and to allow the use of more aggressive disinfection treatments.