Laboratory of Cell Culture Technology and Biotransformation, Faculty of Food Technology and Biotechnology, University of Zagreb, 6 Pierotti St., 10000, Zagreb, Croatia.
Cytotechnology. 2010 Dec;62(6):585-94. doi: 10.1007/s10616-010-9312-y. Epub 2010 Nov 11.
In this study the effects of ammonium and lactate on a culture of channel catfish ovary (CCO) cells were examined. We also made investigation on the influence of glutamine, since our previous research revealed that this amino acid stimulated CCO cell growth more than glucose in a concentration-dependent manner. The effect of ammonium in cell culture included the considerable decrease in cell growth rate with eventual growth arrest as well as the retardation of glucose consumption. At ammonium concentrations above 2.5 mM, the cells displayed specific morphological changes. The effect of lactate was different to that of ammonium since the cell growth rate was progressively decreasing with the increase of lactate concentration, whereas the glucose consumption rate remained almost unchanged. Besides that, it was found that lactate was steadily eliminated from the culture medium when its initial concentration was relatively high. The influence of glutamine on CCO cell propagation showed that nutrient requirements of this cell line were mainly dependent on glutamine rather than glucose. The increase in glutamine concentration led to the increase in cell growth rate and consequent ammonia accumulation while the glucose utilization and lactate production were reduced. Without glutamine in culture medium cell growth was arrested. However, the lack of glucose reversed the stimulating effect of glutamine by decreasing cell growth rate and affecting amino acid utilization.
本研究考察了铵和乳酸盐对斑点叉尾鮰卵巢（CCO）细胞培养的影响。我们还研究了谷氨酰胺的影响，因为我们之前的研究表明，这种氨基酸以浓度依赖的方式比葡萄糖更能刺激 CCO 细胞生长。细胞培养中铵的作用包括细胞生长速率显著下降，最终生长停滞，以及葡萄糖消耗的延迟。在 2.5mM 以上的铵浓度下，细胞显示出特定的形态变化。乳酸盐的作用与铵不同，因为随着乳酸盐浓度的增加，细胞生长速率逐渐降低，而葡萄糖消耗速率几乎保持不变。此外，当乳酸盐的初始浓度较高时，发现其从培养基中稳定消除。谷氨酰胺对 CCO 细胞增殖的影响表明，该细胞系的营养需求主要依赖于谷氨酰胺而不是葡萄糖。谷氨酰胺浓度的增加导致细胞生长速率增加，氨积累增加，而葡萄糖利用率和乳酸盐产量减少。在培养基中没有谷氨酰胺的情况下，细胞生长停滞。然而，缺乏葡萄糖通过降低细胞生长速率和影响氨基酸利用来逆转谷氨酰胺的刺激作用。
