Objective: To investigate the effects of Agave sisalana(A. sisalana) extract on Aedes aegypti(Ae. aegypti) primary cell culture.Methods: Cells of Ae. aegypti were exposed to different concentrations of A. sisalana crude extract(0.18–6.00 mg/m L) for 24 h. Then, the cells were labeled with propidium iodide and subjected to fluorescence microscopy to verify cell viability. In addition, nitric oxide production was measured.Results: Results showed that cells exposed to 6 mg/m L of the crude extract presented a greater percentage of death when compared to control(73.8% ± 9.6% vs. 34.6% ± 9.6%).Furthermore, there was an increase in the nitric oxide production in cells exposed to 6 mg/m L of A. sisalana crude extract [(0.81 ± 0.08) mmol/L] compared to control group[(0.41 ± 0.18) mmol/L].Conclusions: The results show that A. sisalana is cytotoxic to Ae. aegypti and may be used as raw material for new eco-friendly and inexpensive insecticides, since sisal industry discards the liquid waste for the extraction of plant fiber. Aegypti were derived from Aedes aegypti (Ae. Aegypti) primary cell culture. Methods: Cells of Ae. Aegypti were exposed to different concentrations of A. sisalana crude extract (0.18-6.00 mg / m L) for 24 h. Then, the cells were labeled with propidium iodide and subjected to fluorescence microscopy to verify cell viability. In addition, nitric oxide production was measured. Results: Results showed that cells exposed to 6 mg / m L of the crude extract presented a greater percentage of death when compared to control (73.8% ± 9.6% vs. 34.6% ± 9.6%.) Furthermore, there was an increase in the nitric oxide production in cells exposed to 6 mg / m L of A (0.81 ± 0.08) mmol / L compared to control group [(0.41 ± 0.18) mmol / L] .Conclusions: The results show that A. sisalana is cytotoxic to Ae. aegypti and may be used as raw material for new eco-friendly and inexpensive insecticides, since sisal industry discards the liquid wast e for the extraction of plant fiber.