Abstract
Ammonia and lactate are the major byproducts from mammalian cells grown in medium containing glutamine and glucose. Both can be toxic to cells, and may limit the productivity of commercial bioreactors. The transient and steady-state responses of hybridoma growth and metabolism to lactate and ammonia pulse and step changes in continuous suspension culture have been examined. No inhibition was observed at 40 mM lactate. Cell growth was inhibited by 5 mM ammonia, but the cells were able to adapt to ammonia concentrations as high as 8.2 mM. Ammonia production decreased and alanine production increased in response to higher ammonia concentrations. Increased ammonia concentrations also inhibited glutamine and oxygen consumption. The specific oxygen consumption rate decreased by an order of magnitude after an ammonia pulse to 18 mM. Under these conditions, over 90% of the estimated ATP production was due to glycolysis and a large fraction of glutamine was converted to lactate.
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Abbreviations
- D d−1 :
-
Dilution rate = (volumetric feed rate) (reactor volume)
- P/0 :
-
Molecules of ATP generated per molecule of NADH oxidized
- q ATP mmol cell−1 d−1 :
-
Specific ATP production rate
- \(q_{{\text{O}}_2 } \) mmol cell−1 h−1 :
-
Specific oxygen uptake rate
- q p mmol cell−1 d−1 :
-
Specific product formation rate
- q s mmol cell−1 d−1 :
-
Specific substrate consumption rate
- Y′ p,s mol/mol:
-
True yield of product from substrate
- Y p,s . mol/mol:
-
Apparent yield of product from substrate = q p /q s
- ala:
-
alanine
- NH3 :
-
ammonia
- gluc:
-
glucose
- glu:
-
glutamate
- gln:
-
glutamine
- lac:
-
lactate
References
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Miller, W.M., Wilke, C.R. & Blanch, H.W. Transient responses of hybridoma cells to lactate and ammonia pulse and step changes in continuous culture. Bioprocess Engineering 3, 113–122 (1988). https://doi.org/10.1007/BF00373474
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DOI: https://doi.org/10.1007/BF00373474