Abstract: Using the “EA3-867” cellulase prepared in our laboratory, viable mesophyll protoplasts were isolated from Solanum nigrum L. The protoplasts grew and divided when cultivated in banging drops and thin liquid layer of NT medium, and calli formed. After transfering the calli on Dudits and MS solid medium (both supplemented with zeatin 1 mg/L, NAA 0.5 mg/L), regenerated plantlets had been obtained. The effects of different inositol amounts in NT medium on the growth of protoplasts were compared. The percentage of 1st and 2nd cell division after 5–8 days culture and the number of cell clusters increased in NT medium containing 250 mg/L inositol.