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Patchouli (Pogostemon cablin Benth.) is an aromatic plant that produces essential oil known as patchouli oil. The current problem on the world patchouli oil is the decrease of patchouli oil production, one of which caused by limited patchouli seedlings on the market. Tissue culture technique can be an alternative way to increase the production of patchouli seedlings. Thus, in this study was conducted to determine the effect of using several concentrations of activated charcoal on the initiation medium and to obtain the most optimal combination of Plant Growth Regulator (PGR) (BAP and NAA concentrations) for the initiation medium of patchouli explants. This study used an experimental method with a three-factor of Completely Randomized Design (CRD) consisting of 100 treatment and 3 times replication. The three factors are BAP with 5 concentration levels, NAA with 5 concentration levels, and activated charcoal with 4 concentrations levels. The observation was made during 35 days after planting by observing the percentage of browning explants and the percentage of explants forming callus parameters. The result of this study showed BAP, NAA, and activated charcoal as PGR gave the same results on the percentage of browning explants. BAP and NAA as PGR also gave the same result on the percentage of explants that forming callus, while activated charcoal showed the different results.