The experiments were conducted to identify several factors affecting isolation and culture of mesophyll protoplasts in Solanum melongena var. fructualbo. Higher viable plotoplasts were obtained, when isolated in 1.5% macerozyme, 0.2% macerozyme, 0.6M mannitol, 0.01M MES, 0.2% BSA containing solution adjusted to pH 6.3 for 4 hours. One hour plasmolysis of the material before digestion of leaf tissue was effective for protoplast yield and viability. The method of washing and purification of crude protoplasts, ESS process with 0.7M mannitol and 0.6M sucrose solution. was the best way to get purified protoplasts with viability. As isolated protoplasts were cultured in 8P-KM medium at a density of $2.5{ imes}10^4/ml$, the cells were enlarged after 3 to 5 days from culture, subsequently the cells were divided and resulted in colonies.