The growth and cell composition of a newly isolated strain of Chlorella pyrenoidosa SHOU-1002 were compared in three mixotrophic groups (treated with glucose, sodium glutamate and yeast extract, respectively), and a photoautotrophic group. The results showed that, generally, the cell density of mixotrophic groups were significantly higher than that of the photoautotrophic group after inoculation. Moreover, the cell density of the glucose group was significantly higher than that of the sodium glutamate group and the yeast extract group within 6 days after inoculation. Additionally, there was no significant difference in the cell density of the three mixotrophic groups from the 10th day. On the 12th day, the contents of chlorophyll a, carotenoids, protein and total lipid in the sodium glutamate group and yeast extract group were significantly higher as compared to those in the glucose group; however, the carbohydrate content was significantly lower compared to the glucose group. Further, in the glucose group, SFAs increased while the PUFAs decreased with the prolongation of culture time. The PUFAs in the sodium glutamate group and the yeast extract group tended to decrease in the beginning, then they increased. The PUFAs of the sodium glutamate group and the yeast extract group were significantly higher than that of the glucose group on the 12th day. In conclusion, the results suggested that Chlorella pyrenoidosa SHOU-1002 had the ability of mixotrophy and its growth efficiency was significantly improved in mixotrophic mode. Besides, the algal cells cultured by yeast extract or sodium glutamate had higher nutritional value than those cultured by glucose.