이상에서 살펴본 바와 같이 하이브리도마 세포의 고농도 배양에서 ammonia 가 세포 생존도에 미치는 영향을 조사하고 또 그 기작은 어떠한 것인가를 알아 보았다. 또한 세포내 ammonia 농도와 세포내 ATP 농도는 세포 생존도와 밀접한 관계를 갖고 있음을 알 수 있다. 하이브리도마 세포의 고농도 배양에서 높은 세포 생존도를 유지하기 위해서는 배양액 내의 ...
이상에서 살펴본 바와 같이 하이브리도마 세포의 고농도 배양에서 ammonia 가 세포 생존도에 미치는 영향을 조사하고 또 그 기작은 어떠한 것인가를 알아 보았다. 또한 세포내 ammonia 농도와 세포내 ATP 농도는 세포 생존도와 밀접한 관계를 갖고 있음을 알 수 있다. 하이브리도마 세포의 고농도 배양에서 높은 세포 생존도를 유지하기 위해서는 배양액 내의 ammonia 농도나 glucose의 농도를 적절히 조절하는 것이 필요하며 이를 위해서는 배양액 내 이들의 농도를 연속적으로 측정하는 방법이 개발되어야 할 것이다.
이상에서 살펴본 바와 같이 하이브리도마 세포의 고농도 배양에서 ammonia 가 세포 생존도에 미치는 영향을 조사하고 또 그 기작은 어떠한 것인가를 알아 보았다. 또한 세포내 ammonia 농도와 세포내 ATP 농도는 세포 생존도와 밀접한 관계를 갖고 있음을 알 수 있다. 하이브리도마 세포의 고농도 배양에서 높은 세포 생존도를 유지하기 위해서는 배양액 내의 ammonia 농도나 glucose의 농도를 적절히 조절하는 것이 필요하며 이를 위해서는 배양액 내 이들의 농도를 연속적으로 측정하는 방법이 개발되어야 할 것이다.
In high cell-density hybridoma culture for monoclonal antibody production using perfusion culture system, sudden cell deaths were observed as the cell concentration increased. Sudden cell deaths can be attributed to the various reasons such as the nutrient depletion in a culture medium, accumulation...
In high cell-density hybridoma culture for monoclonal antibody production using perfusion culture system, sudden cell deaths were observed as the cell concentration increased. Sudden cell deaths can be attributed to the various reasons such as the nutrient depletion in a culture medium, accumulation of toxic waste, changes in physicochemical milieu, cellular membrane damage, and membrane destruction by intracellular enzymes. In this study, the relationship between ammonia toxicity and cell viability was investigated. In the initial culture medium, ammonia concentration affected cell growth to some extent from that of above 35㎍/ml. After thirty hours of culture time, ammonia concentration even reaching to that of 90㎍/ml did not have great influence on cellular growth. In addition, influences of ammonia concentration on cell growth in each culture medium varied even under the same concentration. This phenomenon implies that ammonia concentration in a culture medium alone cannot represent the ammonia as a whole actually affecting cell viability. Ammonia concentration in a culture medium differed in its inhibition effect according to the culture tire and its effect, on the contrary, decreased gradually in higher concentrations. At the initial high ammonia concentration, the intracellular production rate of ammonia is slowed down due to the depressed cellular growth rate, resulting in the decreased inhibition effect as the culture time lengthens. The influences of ammonia concentration on cell growth inhibition and cell viability were also found to be different according to the culture method, for two reasons. First, the ammonia concentration in a culture medium is net always proportional to the intracellular concentration. Secondly, its influence on cells is different according to each culture condition even under the same concentrations. Intracellular ammonia concentration decreased at the initial lag phase and increased at the exponential phase and also increased as the cell viability decreased. In the relationship between the cell viability and intracellular ATP concentration, marked decrease in cell viability was observed at the intracellular ATP concentration below 3PN/cell. Though ammonia concentration in a culture medium is not necessarily proportional to the intracellular one, proportional relationship was observed in this study. The critical ammonia concentration initiating the cell growth inhibition cannot he defined accurately and is assumed to be different even on the same cell line according to the cell culture time, the nutrient condition in a culture medium and the cell concentration. Therefore, it is critical in high cell-density hybridoma culture to maintain appropriate ammonia and glucose concentrations in a culture medium for higher cell viability.
In high cell-density hybridoma culture for monoclonal antibody production using perfusion culture system, sudden cell deaths were observed as the cell concentration increased. Sudden cell deaths can be attributed to the various reasons such as the nutrient depletion in a culture medium, accumulation of toxic waste, changes in physicochemical milieu, cellular membrane damage, and membrane destruction by intracellular enzymes. In this study, the relationship between ammonia toxicity and cell viability was investigated. In the initial culture medium, ammonia concentration affected cell growth to some extent from that of above 35㎍/ml. After thirty hours of culture time, ammonia concentration even reaching to that of 90㎍/ml did not have great influence on cellular growth. In addition, influences of ammonia concentration on cell growth in each culture medium varied even under the same concentration. This phenomenon implies that ammonia concentration in a culture medium alone cannot represent the ammonia as a whole actually affecting cell viability. Ammonia concentration in a culture medium differed in its inhibition effect according to the culture tire and its effect, on the contrary, decreased gradually in higher concentrations. At the initial high ammonia concentration, the intracellular production rate of ammonia is slowed down due to the depressed cellular growth rate, resulting in the decreased inhibition effect as the culture time lengthens. The influences of ammonia concentration on cell growth inhibition and cell viability were also found to be different according to the culture method, for two reasons. First, the ammonia concentration in a culture medium is net always proportional to the intracellular concentration. Secondly, its influence on cells is different according to each culture condition even under the same concentrations. Intracellular ammonia concentration decreased at the initial lag phase and increased at the exponential phase and also increased as the cell viability decreased. In the relationship between the cell viability and intracellular ATP concentration, marked decrease in cell viability was observed at the intracellular ATP concentration below 3PN/cell. Though ammonia concentration in a culture medium is not necessarily proportional to the intracellular one, proportional relationship was observed in this study. The critical ammonia concentration initiating the cell growth inhibition cannot he defined accurately and is assumed to be different even on the same cell line according to the cell culture time, the nutrient condition in a culture medium and the cell concentration. Therefore, it is critical in high cell-density hybridoma culture to maintain appropriate ammonia and glucose concentrations in a culture medium for higher cell viability.
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