Objective: Despite advances in revascularization therapy, coronary artery disease (CAD) poising a leading cause of morbidity and mortality risks worldwide, continues to challenge the medical community. This study aimed at examining the efficacy of bone marrow stem cells combination therapy used for cardiovascular regenerative therapies, and also to identify isolation procedure of autologous bone marrow stem cells (ABMSCs) might affect cell functionality and therapeutic efficacy, administrated through intra-myocardial and during coronary artery bypass graft (CABG).Materials and Methods: The Yazd Cardiovascular research center was investigating to establish a xeno-free protocol with lees processing steps for cell therapy and treatment of cardiovascular disease. After thoughtful consideration of both statistical and clinical principles, we recruited 15 patients to receive either combinations of ABMSCs or placebo. The overall baseline age of the patients was 60+9 years, and due to severe ischemic cardiomyopathy, need to have coronary bypass surgery. Each patient received a median of 5×107±1×107 ABMSC combinations harvested by the special procedure or an equal volume of diluent (placebo). Changes in left ventricular ejection fraction from baseline and wall motion, examined by magnetic resonance imaging and by echocardiography, respectively.Results: ABMSC delivery was successful in all patients with no arrhythmias, no elevated cardiac enzymes or complications related to the delivery. Compared with the placebo-control group, the ABMSC group had significant changes in routine clinical and biochemical test values (p<0.05). Weekly angina frequency was significantly lower in the cell-injected group than in placebo group. Left ventricular ejection fraction, end-systolic volume and wall motion was found to be significantly improved in the ABMSC group compared to the placebo group over the follow-up period.Conclusion: The designed protocol should be considered as an alternative method of ABMSC application in clinical trial for improving blood flow and contractile function of the heart, and also to safely deliver large number of cells regardless of different stem cell processing steps and RBC or xeno-contamination. This study indicates that the beneficial effect of special way which we used for ABMSC transplantation, would be pronounced in CAD patients.