Growth behavior of Chinese hamster ovary cells in a compact loop bioreactor. 2. Effects of medium components and waste products.
Effects of biochemical factors, i.e., medium components and metabolic byproducts, on growth of Chinese hamster ovary (CHO) cells were investigated. Glucose and ammonia were found to inhibit the growth. Kinetic analysis gave the inhibition constants, 0.14 g l-1 for ammonia and 5.0 g l-1 for glucose. Since glutamine was unstable and was a main source of ammonia, precise studies on glutamine degradation and ammonia formation process were done. By evaluating the spontaneous reactions, net glutamine utilization and net ammonia production by the cells could be estimated. It became evident that asparagine could support the growth of CHO cells as a stable substitute for glutamine. Then, a glucose fed-batch culture was grown on a glutamine free and asparagine supplemented medium. Because of (1) low glucose concentration, but (2) no glucose limitation and (3) low ammonia accumulation, the maximum total cell concentration reached 3.4 x 10(6) ml-1, which was 1.8 times greater than that in the control experiment (initial 1.15 g l-1 glucose and 0.29 g l-1 glutamine, and no glucose feed).
Department of Biotechnology, Swiss Federal Institute of Technology, Zürich.
N
N
Kurano
C
C
Leist
F
F
Messi
S
S
Kurano
A
A
Fiechter
D000641:Ammonia / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D001216:Asparagine / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects*; D002460:Cell Line; D002478:Cells, Cultured / Q000166:cytology*; D006224:Cricetinae; D005260:Female; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology*; D005973:Glutamine / Q000378:metabolism; D007700:Kinetics; D007773:Lactates / Q000378:metabolism; D019344:Lactic Acid; D010053:Ovary; D010084:Oxidation-Reduction
D007773:Lactates; D005973:Glutamine; D019344:Lactic Acid; D001216:Asparagine; D000641:Ammonia; D005947:Glucose