Post-mortem fertilization is considered a technique in which semen is extracted from a dead male to fertilize the egg of a living female. Therefore, this technique can be an excellent option in the face of the loss of biodiversity of multiple animal species. The objective of this research is to implement post-mortem assisted reproduction in the charal, Chirostoma jordani, which is a species of economic, biological and cultural importance in Mexico. Mature males were obtained from commercial capture Atlangatepec Dam, Tlaxcala, Mexico. The mature males were sacrificed and divided into three groups: 1) room temperature 18±1°C, 2) ice 5±1°C and 3) refrigeration 11±1°C. The variables evaluated were: percentage (s), duration of motility (%), and fertilization rate (%) at zero, two, four, six, eight and ten hours in quadruplicate, respectively. The results indicate that sperm viability decreased over time; at room temperature it lasted six hours; in ice and refrigeration 10, denoting significant differences (P<0.05). Semen from organisms preserved in ice and refrigeration presented the best fertilization percentages at two and four hours (>90 and ~80%, respectively) (P<0.05). For practical purposes, it was demonstrated that semen from post-mortem organisms can be used to develop conservation practices.