(1) Background: Segmentation of the bladder inner’s wall and outer boundaries on Magnetic Resonance Images (MRI) is a crucial step for the diagnosis and the characterization of the bladder state and function. This paper proposes an optimized system for the segmentation and the classification of the bladder wall. (2) Methods: For each image of our data set, the region of interest corresponding to the bladder wall was extracted using LevelSet contour-based segmentation. Several features were computed from the extracted wall on T2 MRI images. After an automatic selection of the sub-vector containing most discriminant features, two supervised learning algorithms were tested using a bio-inspired optimization algorithm. (3) Results: The proposed system based on the improved LevelSet algorithm proved its efficiency in bladder wall segmentation. Experiments also showed that Support Vector Machine (SVM) classifier, optimized by Gray Wolf Optimizer (GWO) and using Radial Basis Function (RBF) kernel outperforms the Random Forest classification algorithm with a set of selected features. (4) Conclusions: A computer-aided optimized system based on segmentation and characterization, of bladder wall on MRI images for classification purposes is proposed. It can significantly be helpful for radiologists as a part of spina bifida study.