The all-trans-retinoic acid (ATRA) is the most active form of vitamin A that helps to regulate the proliferation, differentiation and apoptosis of several types of cells, mainly the adipocytes, and causes weight loss through the reduction of adipogenesis and lipogenesis. In this present study we demonstrated that ATRA concentrations of 20.75, 50 and 100 μM decreased the cell viability in vitro of human adipose-derived stem cells (ADSCs), and in ADSCs during adipogenic differentiation. The cells cycle assessment showed that ATRA increased the cell frequency in Sub-G1 at 4.02x and decreased it in G1 in 2.54x. Moreover, the membrane integrity loss increased by 4.66x and apoptosis increased by 33.56x in ATRA-treated cultures. The gene expression assay suggested that the treatment using ATRA leads to mitochondrial membrane permeabilization and to consequent release of proapoptotic BAK and BAX molecules (increased expression 5.5 and 5.4x respectively); in addition, it increased CASP3 expression (by 8.8x). These events may activate the Bcl-2 (4.1x increase), GADD45 (increase 3.14x) and PPAR-γ (16x increase) expressions, as well as, to reduce the p53 (by −1.38x) expression; therefore, these events should be further mediated by increased RARα expression (by 3.8x). The results evidenced that ATRA may be a good proposal for mesotherapy strategies in order to control the development of subcutaneous adipose tissue; as this tissue have a higher development in some specific areas and ATRA interferes not only in the ADSCs differentiation but also in the apoptosis of ADSCs, preadipocytes and adipocytes.