Under glutamine-limited condition, $2 imes10^6$ (viable cells/ml) of maximum cell density and 13.5 ($mu g$/ml) of tissue-type Plasminogen Activators (tPA) production were maintained by spike feeding fresh medium in fed-batch cultivation of human recombinant melanoma cells. It showed that tPA production was much seriously affected than cell growth according to initial glutamine concentrations. Above 3.4 (mmol/I) of glutamine concentration both cell growth and tPA production were not much affected by increasing initial glutamine concentration. Glutamine depleted situation was occurred at latter periods of batch and fed-batch cultivations below 5.4 (mmole/I) of initial glutamine concentration. It also showed that maximum glutamine consumption and ammonia evolution rates were closely related to initial glutamine concentrations. Maximum specific tPA production rate was estimated as $8.1 imes19^{-6}$ ($mu g$/cells/h) at 3.4(mmol/I) of glutamine concentration, which is higher than that from other batch and fed-batch processes.