The study aims to determine the genetic variability among eleven somaclones of sugarcane obtained via immature leaves of the Egyptian commercial variety GT-54 9 at Agronomy Department, Faculty of Agriculture, Assiut University.  Significant differences were found among the somaclones and also their donor, revealing to abundance genetic differences among them. The highest values of GCV and PCV were estimated in agronomic traits for cane yield (18.11 and 18.53%) and in technological traits for sugar yield (17.65 and 17.76%) over two ratoon crops, respectively. The heritability in agronomic traits ranged from 50.39 (stalk diameter) to 98.46% (cane yield) and in technological traits varied from 73.02 (purity) to 98.78% (sugar yield) over the both ratoon crops. The obtained significant differences coupled with GCV, PCV and heritability estimates explained the differences among the studied somaclones (somaclonal variation). The average over the two ratoon crops revealed that somaclones no. 7 and 8 surpassed the donor in highly significant values for most agronomic traits i.e. stalk height (14.35 and 9.48%), stalk weight (9.52 and 15.24%), stalk number/fed (21.00 and 31.25%) and cane yield (32.16 and 52.02%), respectively. Also, the somaclone no. 4 surpassed the donor in highly significant values for all technological traits i.e. sugar yield (23.52), brix (3.13), sucrose% (6.28), purity% (2.99), pol% (6.36) and sugar recovery% (7.72%). The results concluded that the In-vitro cultures exhibited genetic variability among the obtained somaclones (somaclonal variation), which could be used to invent new superior somaclones and overcome the accomplishments of traditional cane breeding.