In Ecuador, banana production is being threatened by the age of plantations and diseases like Mycosphaerella fijiensis Morelet. Although bananas are traditionally propagated by suckers, by using tissue-cultured plants it is possible to produce larger quantities of healthy plantlets for both small- and large-scale farmers. The aim of this study was to develop alternative methods for the micropropagation of banana cv. Williams, as well as for the acclimation of the obtained plantlets. We present three sets of experiments i) in vitro establishment of genotypes of banana growing at “El Oro” (Ecuador), ii) micropropagation of banana in liquid medium by temporary immersion, and iii) use of extracts of Moringa oleifera Lam as a biofortificant during the acclimation of banana vitroplants. The obtained results indicate: i) the need to follow extremely careful sterilization protocols for establishing banana cultures, in order to counteract the presence of fungal and bacterial contaminants due to the climate conditions of “El Oro” (Ecuador), ii) the potential of temporary immersion systems for propagation of banana, and iii) the beneficial effects of extracts of Moringa leaves on vitroplants of banana during the acclimation step.