Tinospora cordifolia, a medicinal plant known for its rich bioactive compounds, holds significant pharmacological importance due to its alkaloid and phenolic content. In this study, the effect of gibberellic acid (GA), a plant growth regulator, on the enhancement of total phenols and alkaloids in in vitro cell cultures of T. cordifolia was investigated. Cells were treated with different concentrations of GA (1–5 mg/L), and the maximum metabolite production was observed at 3 mg/L after 30 days of culture. GA treatment led to a significant increase in total alkaloid content, peaking at 9.454 mg/g DW in the cell extracts, demonstrating GA’s role in stimulating alkaloid biosynthesis. Similarly, total phenol content was highest at 3 mg/L GA, with 20.5 mg/g DW in the cell extracts, confirming GA’s influence on phenolic pathways. These results highlight GA’s potential as an elicitor for enhancing the production of bioactive compounds, offering promising applications for large- scale secondary metabolite production in Tinospora cordifolia under controlled conditions.