Introduction: Adult stem cells (ASCs) are a population of tissue-resident cells that have the capacity for self-renewal and differentiation into different cell types with potential for cell therapies. New approaches for ASCs isolation, including many tissue sources and new protocols that are more effective and less expensive are under investigation. Thus this work aim is to isolate, maintain in cell culture and evaluate cryopreservation protocols for adipose derived stem cells (ADSCs) from different tissues such as subcutaneous adipose tissue, visceral mesenteric and omental visceral taken from the same individual. Material and Methods: The techniques of mechanical and enzymatic dissociation were used, in order to investigate the most appropriated method to ADSCs isolation. Dimethylsulfoxide (DMSO) and dimethylformamide (DMF) in different concentrations were tested as cryoprotectors in 24, 48 and 72 hours thawing protocols. The samples were collected from obese patients with associated diseases undergoing bariatric surgery, between 30 to 45 years old. Results: Among 10 collected samples it was possible to measure cell viability from 4 patients. The higher cell rate was obtained from the visceral tissue of omentum. Conclusion: DMSO was the more efficient cryopreservant for this condition. This adipose tissue source could be explored for ADSCs isolation and future clinical investigations.