Proper selection and application of preprocessing steps are crucial for obtaining accurate segmentation in brain Magnetic Resonance Imaging (MRI). The aim of this study is to evaluate the impact brain extraction (BE) and bias field correction (BFC) methods have on regional brain volume (RBV) measurements of preterm neonates’ T2w MRI at term-equivalent age (TEA). Five BE methods (Manual, BET2, SWS, HD-BET, SynthStrip) were applied together with two BFC methods (SPM-BFC and N4ITK), before segmenting the neonatal brain into eight tissue classes (cortical grey matter, white matter, cerebral spinal fluid, deep nuclear grey matter, hippocampus, amygdala, cerebellum, and brainstem) using an automated segmentation software (MANTiS). Quantitative assessments were conducted, including the coefficient of variation (CV), coefficient of joint variation (CJV), Dice coefficient (DC), and RBV. HD-BET, together with N4ITK, showed the highest performance (mean ± standard deviation) regarding CV of 0.047 ± 0.005 (white matter) and 0.070 ± 0.005 (grey matter), CJV of 0.662 ± 0.095, DC of 0.942 ± 0.063, and RBV without significant differences (except in the brainstem) from the manual segmentation. Therefore, such combination of methods is recommended for improved skull-stripping accuracy, intensity homogeneity, and reproducibility of RBV of T2w MRI at TEA.