Abstract: The effects of carbon and nitrogen sources as well as carbon-to-nitrogen mass ratio on the growth of Chlorella sp USTB01 and the production of lutein were investigated under the condition of heterotrophic culture in.dark. The results show that glucose and potassium nitrate were the best carbon and nitrogen sources to support the rapid and continuous growth of Chlorella sp USTB01 respectively. When glucose and potassium nitrate were used as the sole carbon and nitrogen sources and the initial concentration of nitrogen was 0.28 g·L^-1, the growth of Chlorella sp increased with the increase of carbon-to-nitrogen mass ratio in the range from 15 to 25. Potassium nitrate was the best nitrogen source to promote biosynthesis of lutein by Chlorella sp USTB01 and the content of lutein in the cells of Chlorella sp USTB01 declined with the increase of carbon-nitrogen mass ratio from 15 to 30.