Abstract: Background and Objective: Solanum surattense (S. surattense) is an important medicinal plant used for several medicinal purposes. Seaweed extract is being used as a biostimulant due to the presence of plant growth regulators. Hence in this study, seaweed extract was used in combination with cytokinins to develop an efficient protocol for in vitro regeneration of Solanum surattense. Methodology: The explants were excised from 7 days old seedling and inoculated in MS medium supplemented with different concentrations and combinations of cytokinins and seaweed extract. Data were analyzed using SPSS software version 17 by one-way ANOVA. Results: The best number of shoots (8.5) with 100% of response was observed in cotyledonary node at 1.5 mg L–1 6-Benzylaminopurine (BAP)+30% Turbinaria decurrens extract (TDE) and 80% of explants have responded on 1.5 mg L–1 of BAP alone. The excised shoots from the multiple shoots were cultured on MS medium supplemented with Indole-3-butyric acid (IBA) at 0.4 mg L–1 of concentration produced roots (5.6) with 89.6% of response was observed. The excised shoots were cultured on MS media fortified with the combination of IBA 0.4 mg L–1 and 20% of Turbinaria extract produced maximum number of roots (6.2)/shoot with 95.6% of response was found. Conclusion: The seaweed extract and plant growth regulators are important to promote the Solanum surattense plant regeneration and also noted that the cotyledonary node explant is the suitable material for the production of more number of multiple shoots.