In this study, we performed an adipogenic differentiation of human adipose-derived stem cells (ADSCs) in vitro with different deuterium content (natural, low and high) in the culture medium during differentiation process with parallel analysis of the gene expression, metabolic activity and cell viability/ toxicity. After ADSCs differentiation into adipocytes we have done the analysis of differentiation process efficiency and determined a type of resulting adipocytes (by morphology, gene expression, UCP1 protein detection and adipokine production analysis). We have found that high (5 × 10 5 ppm) deuterium content significantly inhibit in vitro adipogenic differentiation of human ADSCs compared to the groups with natural (150 ppm) and low (30 ppm) deuterium content. Importantly, protocol of differentiation used in our study leads to white adipocytes development in groups with natural (control) and high deuterium content, whereas deuterium-depleted differentiation medium leads to brown-like (beige) adipocytes formation. We have also remarked the direct impact of deuterium on the cellular survival and metabolic activity. Interesting, in deuterium depleted-medium, the cells had normal survival rate and high metabolic activity, whereas the inhibitory effect of deuterated medium on ADSCs differentiation at least was partly associated with deuterium cytotoxicity and inhibitory effect on metabolic activity. The inhibitory effect of deuterium on metabolic activity and the subsequent decrease in the effectiveness of adipogenic differentiation is probably associated with mitochondrial dysfunction. Thus, deuterium could be considered as an element that affects the substance chirality. These findings may be the basis for the development of new approaches in the treatment of obesity, metabolic syndrome and diabetes through the regulation of adipose-derived stem cell differentiation and adipocyte functions. In the 21st century, non-communicable diseases (NCD) like obesity, metabolic syndrome and type 2 diabetes mellitus (T2DM) became the main medical problems of the humanity 1-4. These diseases started in the Western world, but in parallel with the improving of human life standards, technological progress and the spread of the Western lifestyle also around the world. So, these diseases have become a global epidemic 5. Currently, although there remains a correlation between the level of economic development and the frequency of these diseases, they have ceased to be a medical problem in high-income countries, but also have become an urgent item for the low-income and middle-income countries 6. A characteristic feature of obesity, metabolic syndrome and T2DM is the defection of glucose and lipids metabolism, which is manifested in insulin resistance, impaired fasting glucose, dyslipidemia, high blood sugar, high serum triglycerides, imbalance of different types of lipoproteins in blood serum 7,8 .