Many endangered freshwater turtle species are born in captivity for conservation and future reintroduction into the wild. However, in order to improve breeding programs, an assessment of the genetic diversity of the founder individuals is required to avoid genetic problems such as inbreeding, fixation of deleterious alleles, or loss of allelic diversity due to genetic drift. In this research, we assessed the genetic diversity of the founder individuals from three Wildlife Management Units (UMA) dedicated to the reproduction of Dermatemys mawii in southeast Mexico, and from three wild populations using ten microsatellite markers. Dermatemys mawii is a freshwater turtle that is critically endangered due principally to fragmentation, loss, degradation, and contamination of its habitat, in addition to hunting for human consumption. Furthermore, genetic relationships among UMAs and wild populations, as well as within each kind of group, were investigated by means of Bayesian analysis (Structure software) and discriminant analysis of principal component (DAPC). Genetic diversity in wild populations could be considered as medium, and are less than values observed for UMAs. Genetic diversity for UMAs and wild populations were discussed considering origin of individuals, translocation between UMAs, habitat quality among other factors. Genetic structure analysis highlighted an evident separation between UMAs and wild populations (Bayesian and DAPC analyses), and the hierarchical analysis of structure among UMAs reflected the origin and relationship among them, whereas geographical situation of wild populations is the best explanation for its hierarchical structure. In light of our results, some conservation and management recommendations are provided for this endangered freshwater turtle.