Na-butyrate sustains energetic states of metabolism and t-PA productivity of CHO cells

Abstract

The effects of Na-butyrate on the physiological behaviour and on the specific productivity of recombinant tissue plasminogen activator (t-PA) Chinese Hamster Ovary (CHO) cells were characterized. Batch cultures were performed in a 3.5-L bioreactor. Na-butyrate was added either at the mid-exponential growth phase (48 h) or at the end of the exponential growth phase (74 h). The cultures with Na-butyrate showed higher net specific productivity of t-PA and lower final cell density and viability. Maximum specific productivity of t-PA for all cultures coincided with the early plateau phase (84 h). The cell's specific oxygen uptake rate (qO2) increased after the Na-butyrate addition and remained higher than that of the controlled culture. Triphosphate nucleotides, ADP, AMP and UDP-sugars all increased after 84 h in the cultures with Na-butyrate, showing different behaviours when Na-butyrate was added at 48 h or 74 h. Na-butyrate did not affect the cell's adenylate energy charge until the cell's viability started to decrease (156-168 h). The controlled culture and the culture with Na-butyrate addition, showed at 74 h, similar time trends as for purine and nucleotide ratios ((ATP+GTP)/(UTP+CTP) and UTP/ATP) with clear shifts in behaviour at 84 h and 168 h. However, the addition of Na-butyrate at 48 h resulted in damped variations of purine and nucleotide ratios in comparison to both the control culture and the culture with Na-butyrate addition at 74 h.