Jerusalem artichoke (JA) family (Asterceae) is a perennial species known for its tubers rich in inulin, a valuable source of fructose for diabetics.This study aimed to establish an applicable protocol for calli induction and production from different explants of Jerusalem artichoke (Balady, Fuza and Alba) cultivars. For this purpose; leaf, stem and root explants derived from in vitro growing plantlets were cultured on Murashige and Skoog (MS medium) augmented with different combinations of benzyl adenine (BA) and naphthalenacetic acid (NAA).The highest percentage of calli induction (100%) was recorded with all tested media except MS free growth regulators medium and MS fortified with 1mg L-1 BA. On the other hand, the maximum value of calli fresh weight was obtained by culturing stem explants on MS supplemented with 1 mg L-1 BA + 2 mg L-1 NAA for all cultivars. NAA for all cultivars. Moreover and concerning calli growth dynamic for each cultivar, the recorded data clearly showed that gradually increase in calli fresh weights by week reaching the maximum value at eighth week for all used explants cultured on the best medium for calli initiation. The results proved that Balady cultivar recorded the best results for calli induction and production compared with Fuza and Alba cultivars. In the same context, stem explant was the superior explant when it compared with root and leaf explants for all examined cultivars. Moreover and concerning calli growth dynamic for each cultivar, the recorded data clearly showed that gradually increase in calli fresh weights by week reaching the maximum value at eighth week for all used explants cultured on the best medium for calli initiation. The results proved that Balady cultivar recorded the best results for calli induction and production compared with Fuza and Alba cultivars. In the same context, stem explant was the superior explant when it compared with root and leaf explants for all examined cultivars. MS medium fortified with 1 mg L-1 BA + 2 mg L-1 NAA was the most suitable medium compared to the other tested media for inducing calli and to enhance calli production from stem explant for all cultivars.