Abstract: To investigate the improvement effects of stem pre-pressing forward shift on filament structure of tobacco stems from different origins, and to clarify differentiated pre-pressing process paths and key control indicators, tobacco stems from four producing areas (Fujian, Yunnan, Henan, and Jilin) were used as experimental materials. Three pre-pressing paths were designed: single pre-pressing with a roll gap at 0.8 mm (T1), two-stage continuous pre-pressing with roll gaps at 0.8 mm and 0.4 mm (T2), and single pre-pressing with a roll gap at 0.4 mm (T3). The morphological evolution patterns of stem sheets throughout the entire process of pre-pressing, drying, and re-pressing, as well as their correlation with stem filament structure, were systematically analyzed. The results showed that: 1) Pre-pressing treatment significantly improved the quality of stem filament structure. The average width of stem filaments from the four areas decreased by 0.20 mm–0.28 mm under three paths, and the proportion of filamentous stem filaments increased by an average of 15.4–22.0 percentage points. 2) Tobacco stem tissue structure significantly influenced the response to pre-pressing paths. Stems with dense structures, such as those from Fujian and Yunnan, exhibited better filament structure under T3 path, while stems with loose and porous structures, such as those from Henan, showed lower sensitivity to pre-pressing paths. Stems from Jilin fell between these two types. 3) The thickness of pre-pressed stem sheets was a source indicator affecting stem filament width, and repeated pressing with the same gap could not further improve filament formation. For the implementation of tobacco stem pre-pressing processing, it is recommended to use the thickness of non-vascular tissue in original stems as a predictive basis for differentiated process path and parameter settings, with pre-pressed stem sheet thickness as the key control indicator and dried stem sheet width as the process monitoring indicator.