Introduction: Bacopa Monnieri, commonly known, as “Brahmi” is a medicinal herb extensively used in Ayurveda for its neuro protective properties, including memory enhancement, stress reduction, and potential therapeutic effects against neurodegen erative diseases such as Alzheimer’s’. Due to its increasing demand, an efficient micropropagation protocol is necessary for large-scale and sustainable production. Aim/Objective: This study aims to develop and optimize an in vitro micropropagation protocol for Bacopa Monnieri using shoot explants cultured on Murashige and Skoog (MS) medium supplemented with different plant growth regulators (PGRs). Methods: Shoot explants were cultured on MS medium with varying concentrations of Indole-3-acetic acid (IAA), 6-Benzylami nopurine (BAP), Kinetin (Kn), Naphthalene acetic acid (NAA), and 2,4-Dichlorophenoxyacetic acid (2,4-D). The cultures were evaluated for shoot initiation, elongation, and multiplication over a 30-day period. Gamborg’s B5 medium was also tested for comparison. Results: Shoot initiation was observed within six days of incubation, with significant growth by the third week. The highest shoot multiplication rate was obtained using MS medium supplemented with 2.5 mg/L BAP + 2.5 mg/L Kn + 0.5 mg/L NAA + 0.5 mg/L 2,4-D. This combination resulted in a 98% response rate, the shortest shoot initiation time (6 days), the highest number of shoots per explant (10.1), and the maximum shoot length (7.9 cm). MS medium demonstrated superior shoot proliferation compared to Gamborg’s B5 medium. Conclusion: The optimized micropropagation protocol facilitates the large-scale production of Bacopa Monnieri, ensuring a sustainable supply for medicinal and commercial applications. This method also contributes to species conservation. Future research should focus on genetic stability, secondary metabolite production, and field trials to validate large-scale cultivation. Key Words: Bacopa Monnieri, Micropropagation, Ayurvedic medicine, Cognitive enhancement, Neuroprotection, Pharmacologi cal properties