In human brain, Corpus Callosum (CC) is the largest white matter structure, connecting between right and left hemispheres. Structural features such as shape and size of CC in midsagittal plane are of great significance for analyzing various neurological diseases, for example Alzheimer’s disease, autism and epilepsy. For quantitative and qualitative studies of CC in brain MR images, robust segmentation of CC is important. In this paper, we present a novel method for CC segmentation. Our approach is based on deep neural networks and the prior information generated from multi-atlas images. Deep neural networks have recently shown good performance in various image processing field. Convolutional neural networks (CNN) have shown outstanding performance for classification and segmentation in medical image fields. We used convolutional neural networks for CC segmentation. Multi-atlas based segmentation model have been widely used in medical image segmentation because atlas has powerful information about the target structure we want to segment, consisting of MR images and corresponding manual segmentation of the target structure. We combined the prior information, such as location and intensity distribution of target structure (i.e. CC), made from multi-atlas images in CNN training process for more improving training. The CNN with prior information showed better segmentation performance than without.