Insulin utilization and kinetic effect on hybridoma metabolism in batch and continuous cultures.
This paper discusses the insulin utilization kinetics and the effect of its concentration during batch and continuous mass cultures of the murine VO208 hybridoma cells, using a home-made serum-free medium. Our results show that insulin is utilized by the cells, with a specific rate of 1 relative units (RU) per 10(9) cells per h in batch culture. In continuous reactor running at different insulin levels, this consumption rate is observed to vary from 0.13 to 0.55 RU per 10(9) cells per h when the insulin activity increases from 0.3 to 35 RU l-1 and then to stabilize for higher insulin levels until 110 RU l-1. A low insulin amount in the medium around 0.01 RU l-1, which is near physiological levels, is found sufficient to promote the cell growth. Interestingly, we observe that too high insulin levels, above 25 RU l-1, induce a reduction of the cell density due to an inhibitory effect on the maximal specific cell growth rate. Furthermore, the specific rate of MAb production is found to be independent of the insulin amount in the medium.
Martial
A
A
Laboratoire des Sciences du Génie Chimique, CNRS-ENSIC, Nancy, France.
Dousset
B
B
Dardenne
M
M
Engasser
J M
JM
Nabet
P
P
Marc
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002455:Cell Division / Q000187:drug effects; D016895:Culture Media, Serum-Free; D046508:Culture Techniques / Q000379:methods*; D004305:Dose-Response Relationship, Drug; D006825:Hybridomas / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology*; D007700:Kinetics; D051379:Mice; D013997:Time Factors
D000911:Antibodies, Monoclonal; D016895:Culture Media, Serum-Free; D007328:Insulin