Objective: Development of science and producing new technology are two important factors which playing main role in inventing new medical treatments for important diseases, like critical heart problems. Heart failure which becomes common in our world, due to increasing the rate of obesity and lacking physical exercises in people lifestyles, is produced by inability of cardiomyocytes in self-renewal. So, using targeted differentiation from mesenchymal multi-potent stem cells into cardiomyocytes could be the sure way for curing this medical condition. Because of this, we focused on finding new pathway for more effective and efficient stem cell targeted differentiation. Materials and Methods: In this experiment, we used Adipose tissue derived stem cells. The extraction method is using collagenase type I on highly fragmented tissue from midscapulare of male rabbit. Flowcytometry used for characterization of extracted cells. Additionally, for examining the multi-potency of these stem cells, the osteogenic and chondrogenicm differentiation have been done. The collagen coated membrane in chemical, mechanical and chemical-mechanical pathways have been used as an appropriate surface for differentiating. The undifferentiated Rabbit stem cells are negative control and cardiomyocytes used as positive one. The interval between affecting stem cells and RNA extraction in our experiments is 4 days. We set GATA 4 (cardiac-specific transcription factor) as target gene besides the HPRT (housekeeping gene) for surveying in Taqman base real time-PCR. Results: The multi-potency of extracted stem cells are approved by the results of osteogenic and chondrogenic differentiation and flocytometry data shows stem cells’ specification. The data from real time-PCR is clearly approved that the mechanotransduction have significant effect on differentiation process by increasing the expression of GATA 4 in treated cells. Conclusion: According to the information that mentioned in results, mechanical loading could be the more appropriate and effective way for targeted differentiation in comparison with chemical factors. Also, the combination of mechanical loading and chemical factors may have more effective influence on adipose derived stem cells for differentiating to cardiomyocyte than other two pathways.