Objective: Mesenchymal stromal cells (MSCs) are multipotent cells with the capacity to differentiate into several cell lineages such as bone, adipose, cardiac and even neural cells in vitro. They are a promising source for cell therapy and tissue engineering nowadays. Today's most culturing media are supplemented with fetal bovine serum (FBS) as the resource of growth factors. But FBS containing culturing media may raise the possibility of zonotic infections and immunological reactions in cell therapy conditions, therefore usage of FBS is considered hazardous for the patients. Numerous researches have been performed to assess the use of FBS-free culturing systems for bone marrow derived mesenchymal stromal cell isolation. Since MSCs can be isolated from several tissues including bone marrow, adipose tissue and umbilical cord blood, we aimed the present investigation to assess the effect of serum free media on growth and differentiate capacity of adipose tissue derived MSCs. Materials and Methods: Approximately 1cm3 surgically waste sterile adipose tissue was digested with collagenase-I leading to a single cell suspension. The isolated cells were cultured in Ultra Culture media supplemented with 2% Ultroser G. MSCs isolation was confirmed with respect to morphology, flowcytometry, adipogenic and osteogenic differentiation potentials. Results: The isolated cells showed adherent spindle-shaped morphology, expanded rapidly and showed expected MSC flowcytometric characteristics; they were positive for CD73, CD90, CD105, CD44, CD166, CD44 and were negative for hematopoietic antigen such as CD45, CD34 and CD14. They could also differentiate successfully into osteoblast and adipocyte which was confirmed with Alizarin Red and Oil red O staining respectively. Conclusion: According to present study, we come to the conclusion that adipose derived MSCs can be cultured in serum-free media with no change in their differentiation capacity. This finding gives us a hope for future cell therapy studies and trials with fewer worries about zonotic infections or immunological reaction.