The chemical decomposition of glutamine is a first-order reaction. Its reaction constants under storage and culture conditions were determined as 0.0009 h-1 and 0.0032 h-1 respectively. Batch culture of hybridoma cell C50 with different initial contents of glutamine helped to understand its real metabolic characteristic. The results show that when the initial concentration of glutamine is lower than 5 mmol/L, more than 80% is used by cells. And the lower the initial content, the more being used. As the initial glutamine concentration increases, the ratio of its utilization decreases. When it reaches 10 nmol/L, the ratio decreases dramatically.