Abstract
The murine myeloma cell line Sp 2/0-Ag 14 was cultured in an ordinary batch culture and in a glutamine limited fed-batch culture. In batch culture, the overflow metabolism of glutamine ends in excess production of ammonium and the amino acids alanine, proline, ornithine, asparagine, glutamate, serine and glycine. This pattern was dramatically changed in the fed-batch culture. Glutamine limitation halved the cellular ammonium production and reduced the ratio of NH4 +/glutamine. The excess production of alanine, proline and ornithine was reduced by a factor of 2–6 while asparagine was not produced at all. In contrary to the other amino acids glycine production was increased. These results are discussed in view of the different nature of glutamine metabolism in the mitochondrial compartment vs. the cytosolic. Furthermore, essential amino acids were used more efficiently in the fed-batch as judged by the increase in the cellular yield coefficients in the range of 1.3–2.6 times for seven of the 11 consumed ones. In all, this leads to a more efficient use of the energy sources glucose and glutamine as revealed by an increase in the cellular yield coefficient for glucose by 70% and for glutamine by 61%.
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Abbreviations
- Nt cells ml−1 :
-
Total cell number
- Nv cells ml−1 :
-
Viable cell number
- Nd cells ml−1 :
-
Dead cell number
- μ h−1 :
-
Specific growth rate
- kd h−1 :
-
Specific death rate
- qp μmol (106 cells h)−1 :
-
Specific product formation rate
- qs μmol (106 cells h)−1 :
-
Specific substrate consumption rate
- YP/S μmol μmol−1 :
-
Yield of product from substrate
- YNt/S 106 cells μmol−1 :
-
Yield of cells from substrate
- YP/Nt μmol (106 cells)−1 :
-
Yield of product from cells
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Ljunggren, J., Häggström, L. Glutamine limited fed-batch culture reduces the overflow metabolism of amino acids in myeloma cells. Cytotechnology 8, 45–56 (1992). https://doi.org/10.1007/BF02540029
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DOI: https://doi.org/10.1007/BF02540029