Growth and metabolism of marine fish Chinook salmon embryo cells: response to lack of glucose and glutamine

Abstract

A peculiar phenomenon, differing from the response of mammalian cells, occurred when Chinook salmon embryo (CHSE) cells were passaged in the medium lacking of both glucose and glutamine. To elucidate metabolic mechanism of CHSE cells, the metabolism parameters, key metabolic enzymes, and ATP levels were measured at different glucose and glutamine concentrations. In the glutamine-free culture, hexokinase activity kept constant, and lactate dehydrogenase (LDH) activity decreased. This indicated that lack of glutamine did not expedite glucose consumption but made it shift to lower lactate production and more efficient energy metabolism. The results coincided with the experimental results of unaltered specific glucose consumption rate and decreased yield coefficients of lactate to glucose. In the glucose-free culture, simultaneous increase of glutaminase activity and of specific ammonia production rate suggested an increased flux into the glutaminolysis pathway, and increases of both glutamate dehydrogenase activity and yield coefficient of ammonia to glutamine showed an increased flux into deamination pathway. However, when glucose and glutamine were both lacking, the specific consumption rates of most of amino acids increased markedly, together with decrease of LDH activity, indicating that pyruvate derived from amino acids, away from lactate production, remedied energy deficiency. When both glucose and glutamine were absent, intracellular ATP contents and the energy charge remained virtually unaltered.

Growth characteristics of channel catfish ovary cells—influence of glucose and glutamine

“…In the medium lacking both glucose and glutamine, CCO cells grew more slowly than cells in complete medium reaching density of 2 9 10 5 cells mL -1 . Cell proliferation in the same medium condition was also reported for chinook salmon embryo (CHSE) cells (Chen et al 2005) and this phenomenon could be specific for some established fish cell lines. The metabolic parameters of glucose and glutamine during eight days of batch culture with different glucose and glutamine concentrations are shown in Table 1.…”

Influence of different ammonium, lactate and glutamine concentrations on CCO cell growth

“…The glutamine to ammonia yield (Y Amm/Gln ) ranged between 1.2 and 1.86 mol 9 mol -1 and was slightly increased in glucose free medium indicating that there was need to use more glutamine as an energy source. The values obtained are in agreement with reports on mammalian and fish cell lines (Cruz et al 1999b;Linz et al 1997;Chen et al 2005). It was to be noted that effects of ammonia accumulation might have been evident in cultures with initial 2 and 4 mM glutamine, regardless of glucose presence in the medium.…”

“…These observations are in agreement with Mancuso et al (1998) and Cruz et al (1999b) who investigated amino acid utilization in hybridomas and BHK cells, respectively. Chen et al (2005) found that in glutamine and glucose free culture of another fish cell line (CHSE) pyruvate was derived from amino acids and covered energy deficiency.…”