The objective of this investigation was to know whether the organophosphate temephos resistance developed in larvae from a laboratory strain of Aedes aegypti (Linnaeus, 1762) from Cuba could be reversed. The resistant laboratory strain of Ae. aegypti, named SAN-F6, was left without temephos selection pressure for 12 generations. The level of temephos resistance was determined using WHO bioassays and mechanisms of metabolic resistance were determined based on enzyme activity levels detected by biochemical assays. Bioassays and biochemical assays were conducted on the SAN-F6 parental strain and every three reversal generations (SANRevF3, SANRevF6, SANRevF9, and SANRevF12) without temephos selection pressure. After 19 yr of keeping the SAN-F6 strain under selection pressure with the LC₉₀ of temephos, the resistance ratio (RR₅₀) was 47.5×. Biochemical assays indicated that esterase and glutathione S-transferase are still responsible for temephos resistance in this strain, but not mixed-function oxidase. Experiments on resistance reversal showed that temephos susceptibility could be recovered as α esterase activity levels decreased. The SAN-F6 strain has provided an essential basis for studies of temephos resistance in Cuba. It was demonstrated that the resistance developed to the larvicide temephos in Ae. aegypti from this Cuban lab strain is a reversible phenomenon, which suggests that similar outcomes might be expected in field populations. As such, the use of temephos alternated with other larvicides recommended by WHO such as Bti or pyriproxyfen is recommended to maintain the effectiveness of temephos and to achieve more effective control of Ae. aegypti.