Abstract: CHO-k1 cells were cultivated in stirred tank bioreactor.Batch culture,fed batch culture and perfusion culture were used,the maximuim viable cell density reached 2.0?106,2.7?106 and 9.6?106mL-1,respectively.The concentration of nutrients (glucose and amino acid) and metabolic by products (ammonia and lactate) were analyzed during cultivation period.The relationship between cellular physiologic condition and nutrient metabolism is discussed in this paper.The energy metabolism in CHO-k1 cells is a typically substrate concentration-dependent overflow metabolism.The limitation of glucose and glutamine decreases by-product formation and increases metabolic efficiency.