Introduction: The productions of generatively mangosteen still has problem because the seeds of mangosteen have recalcitrant characteristic, which means the seeds must be planted immediately, since the seeds does not have dormancy period. Aim: 1) to determine Effect of Plant Growth Regulators on in vitro Callus Induction of mangosteen (Garcinia mangostana L.), 2) to know the best combination and composition of the media to in vitro callus induction of mangosteen (Garcinia mangostana L.). Method: Researcher used complete randomized design (CRD) factorial which is consisting of 12 treatments with 3 times repetitions. The research procedures were: tool sterilization, explant sterilization, media making, explants planting. Result: The fastest response of callus appearance is on sucrose (15; 30 gr / L) and NAA (1,5; 3 ppm) treatment which is producing callus with crumb texture and color is identical to green and white. The callus has 0,53a cm in height, 1,56a cm2 in width, and 0,40a gr in weight. The percentage of callus formation is 100% and the explant response is growing all over the surface (sucrose 15 gr / L + NAA 1,5; 3 ppm). Conclusion: The plant growth regulators have effect on in vitro callus induction of shoot explant mangosteen (Garcinia mangostana L.). The best media combination and composition on in vitro callus induction of shoot explant mangosteen (Garcinia mangostana L.) were sucrose (15; 30 gr / L) and NAA (1,5; 3 ppm).