A callus formation from Aloe vera L. was inducted, and the tissue cultivation of induced callus was investigated. As a result of using a combined growth factors kinetin and NAA (or 2,4-D) varying from 0.5 to 30 μM and 3% sucrose in MS solid medium, the callus grew better than using either kinetin or auxins alone. Supplementation of 2,4-D in the combined growth factor formulation showed an enhanced callus growth. The maximum callus growth occurred when kinetin 5 μM and 2,4-D 30 μM were added in the MS medium. The optimal culture conditions for the callus growth were 25oC, pH 5.5 and 8-10 weeks of cultivation, yielding callus weight of 11.03 mg/aloe tissue (0.5 cm2). Water or ethanol (60%, v/v) extracts from Aloe callus contained the bioactives contents of fresh Aloe vera gel such as glucomannan, and the level of total phenolics content was much higher than that of Aloe vera whole leaf, suggesting the possibility as an alternative production method of Aloe vera bioactives.