Objective To prepare an acute myocardial infarction (AMI) model in Bama mini-pigs by minimally invasive intracoronary balloon occlusion and comprehensively evaluate establishment of the model by hematological, functional, and pathological method. Methods After collecting baseline data and performing a femoral artery puncture under general anesthesia, the left anterior descending branch (2~5 mm below the first diagonal branch) of 12 Bama minipigs was blocked by percutaneous intracoronary balloon occlusion for 90 min. During the operation, Holter monitoring was performed, and intravenous blood was collected regularly after the operation to analyze myocardial injury markers. Echocardiography was also performed after the operation to assess cardiac functions, and the area and severity of myocardial infarction were evaluated by gross observation, and Masson and Sirius red staining. Results Among the 12 mini-pigs, 10 completed the process of balloon occlusion, and the other two did not reach the blocking time because of frequent malignant arrhythmias during the operation. Seven of the animals that completed occlusion survived to 28 days, and the remaining three died unexpectedly before the end point of the experiment. In successfully established models, the ST-T segment of the electrocardiogram showed dynamic changes after occlusion, and myocardial injury markers had increased most obviously at 4 h after the operation compared with before surgery with statistically significant differences (P< 0. 01). Echocardiography on day 7 after the operation showed significant changes in the left ventricular ejection fraction, left ventricular fractional shortening, left ventricular end-diastolic volume, and left ventricular end-systolic volume compared with preoperative values (P< 0. 05). Gross observation of the heart indicated that the infarcted myocardium was gray-white and mainly located in the left anterior wall. Pathological staining showed that the infarct area involved the full thickness of the ventricular wall. Conclusions We established an AMI model in Bama mini-pigs successfully and subsequently conducted comprehensive evaluation of the model validity.