Objective: To explore the application effect of the “dual-line integration teaching mode” in the experimental teaching of systematic anatomy. Methods: A total of 120 students majoring in clinical medicine (Grade 2022) from Hunan University of Medicine were selected and randomly divided into a control group (traditional offline teaching mode) and an experimental group (dual-line integration teaching mode), with 60 students in each group. The control group adopted the traditional teaching mode, where teachers gave explanations and students observed specimens; the experimental group implemented online-offline blended teaching based on the 3D human anatomy specimen teaching system, specimen demonstration videos, and a case-based learning case bank. The teaching effect was evaluated through questionnaire surveys and final comprehensive scores (including theoretical scores, experimental skill assessment scores, and daily performance scores). Results: Compared with the control group, the students in the experimental group showed significant improvements in knowledge mastery ability, hands-on operation ability, teamwork ability, and communication and expression ability (P < 0.05); the final theoretical scores (44.4 ± 3.4 points vs. 40.8 ± 5.3 points) and experimental skill assessment scores (17.4 ± 2.3 points vs. 14.2 ± 3.9 points) of the experimental group were significantly higher than those of the control group, with statistically significant differences (P < 0.05). Conclusion: The “dual-line integration teaching mode” can effectively improve the quality of experimental teaching in systematic anatomy, promote the comprehensive development of students’ learning abilities, and provide new ideas for medical experimental teaching.