To summarize the clinical outcome of totally thoracoscopic cardiac surgery for mitral valve replacement. Clinical data of 634 cases undergoing totally thoracoscopic cardiac surgery for mitral valve replacement from May 2004 to February 2016 in Department of Thoracoscopic Cardiacsurgery, Shanghai Yodak Cardiothoracic Hospital was analyzed retrospectively. There were 292 male and 342 female patients, aged from 17 to 68 years with a mean of (45±13) years. All the 634 patients had moderate-severe mitral valve stenosis and (or) incompetence, 263 patients had moderate-severe tricuspid valve incompetence, 356 patients had atrial fibrillation, 46 patients had left atrium thrombosis. Cardiopulmonary bypass was established with right femoral artery and a single 2 stage venus cannula in the right atrium. The ascending aorta was cross-clamped and the myocardium was protected by coronary perfusion with cold crystalloid cardioplegia. Totally thoracoscopic mitral valve replacement were performed. Thirteen cases had incision expanded and 8 cases had conversions to sternotomy. Cardiopulmonary bypass and aortic cross-clamp time were (89±18) minutes and (51±12) minutes, respectively. Operation time was (3.1±1.2) hours. Mechanical ventilation time and intensive care unit stay were (17±6) hours and (27±8) hours, respectively. Postoperation drainage quantity was (390±70) ml. The hospital days was (9.2±2.1) days. There were 5 cases in-hospital deaths. Postoperative complications occurred in 42 cases (6.6%), including 18 cases of right hemoneumothorax, 12 cases of reoperation for bleeding, 3 cases of perivalvular leakage (reoperation was done in 1 patient), 3 cases of low cardiac output syndrome, 2 cases of acute renal failure, 2 cases of inferior vena cava injury, 1 case of right femoral artery thrombosis and liver injury, respectively. The mean duration of follow-up was (58±9) months in 608 cases, with a follow-up rate of 96.7% (608/629). Three patients had died during the period of follow-up caused by congestive heart failure (2 patients) and stroke (1 patient). Late complication among 605 survivors were 37 cases, including 32 cases of moderate tricuspid valve insufficiency, 3 cases of stroke, 1 case of perivalvular leakage and infective endocarditis, respectively.There was no reoperation during the period of follow-up. Totally thoracoscopic cardiac surgery for mitral valve replacement is safe and effective, with unique superiority and clinical feasible.