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Carbon-13 isotope has a wide range of applications in medical diagnosis, scientific research, food safety and other fields, and the market demand for carbon-13 isotope is also showing an upward trend. In order to explore the separation and production methods of carbon-13 isotope, the gas diffusion separation experiment with ethylene as the processing gas was carried out, and the cascade was designed on this basis. Through the single-stage separation experiment, it can be obtained that the overall separation factor of the gas diffusion method for separating ethylene under the existing experimental conditions is 1.014. The gas diffusion cascade enrichment of carbon-13 isotope involves two steps. The first step is to separate ethylene using a squared-off cascade to obtain the heavy fraction with the carbon-13 isotope abundance reaching 49%. The second step is to convert ethylene into carbon dioxide, and then use a squared-off cascade to separate the carbon dioxide to obtain the heavy fraction with the carbon-13 isotope abundance reaching 95%.