Abstract: Aloe (Aloe vera L., Family Liliaceae), with proven multiple medicinal values finds favour readily in Ayurevdic applications, is facing a serious threat to its population as well as biodiversity due to its popularly harvested aloe leaves by the local communities and herbal medicine vendors. To succeed in dealing with the specified problem along with the facilitation of the mass production of commercial level, constant supply of quality propagules can be possible through conservation of propagules in vitro. The present study is thus concerned with in vitro conservation of multiple shoot culture of aloe to achieve unbroken supply of propagules maintaining their genetic purity. Rhizomatous stem explants result multiple bud break in MS plus 0.25 mg L-1 of NAA and 1.5 mg L-1 of BAP. Separated shoot buds further result in shoot multiplication and proliferation on MS with 2.5 mg L-1 of BAP. In vitro generated multiple shoots were split into individual shoot and subcultured for further multiplication in a sustainable. Five subcultures were performed at an extended 5 month interval over a period of 25 months in the same medium. Plantlets regenerated after 1st subculture and plantlets from 5th subculture showed no significant difference in the phenotypic response. Genetic integrity of in vitro clones was tested using ISSR primers. All monomorphic bands in the ISSR assay, both for primary culture as well long term culture, ascertained their genetic integrity to a great extent.