Abstract: Coccinia grandis (L.) Voigt. (Cucurbitaceae) is an important vegetable with high food and medicinal value. In this plant major problem is seed germination, seed setting and less number of fruit settings. Plant tissue culture technology may help to overcome these problems to a great extent. Therefore, the present study has been designed to develop an efficient protocol for in vitro shoot multiplication, as well as to check its antidiabetic and antioxidant activity. The nodal segments used as explants for cultured. Murashige and Skoog’s (MS) agar-gelled medium with different concentration and combination of BAP, KIN, NAA and NB6 for shoots multiplication and callus. Multiple shoots was seen after 10 days of sub culturing on MS medium. Maximum shoot formed (16±1.5) on MS medium containing 0.5 mg L–1 BAP combination with 0.2 mg L–1 NB6 after 15 days. Antidiabetic activity of C. grandis was also evaluated by using α-amylase inhibition assay and DNS assay. Methanolic extract of fruit, callus (BAP+NB6) and different combination mixture of callus at 1.0 mg L–1 concentration showed 54.29, 75.72 and 80.00% inhibition of α-amylase activity respectively. The result of agar diffusion amylase inhibition assay indicated that methanolic extract of callus show maximum inhibition (46.66%) compare to the fruit extract and fresh callus showed no inhibition. Antioxidant activity of C. grandis was evaluated by using DPPH assay method is based on the reduction of methanolic solution of coloured free radical DPPH by free radical scavenger. The DPPH radical scavenging activity and significantly decreased the formation of oxygen radicals generated in rat peritoneal macrophages plant extracts. The protocol described here is efficient with high number of shoot multiplication and can be employed for the large scale production and genetic manipulation for antidiabetic medicinal plant Coccinia grandis.