Methods for reducing the ammonia in hybridoma cell cultures.
The factors which limit the proliferation of eukaryotic cells in vitro are still not well known. Ammonia is believed to be toxic for mammalian cell proliferation and secretion. We have tried two approaches to reducing the ammonia in the medium. We first limited the ammonia produced by the cells by replacing glutamine by glutamate. Then, we used two chemical engineering methods to eliminate accumulated ammonia. In one the used medium was passed through a natural cation exchanger: the clinoptilolite. In the other, the culture medium was passed through a hydrophobic microporous hollow fiber module. Replacing the glutamine by glutamate reduced the medium ammonia concentration. The physicochemical removal of ammonia induced a better cell growth, but not a better specific antibody secretion.
Capiaumont
J
J
Laboratoire de Biochimie Médicale, Pr Nabet, Faculté de Medecine, Nancy, France.
Legrand
C
C
Carbonell
D
D
Dousset
B
B
Belleville
F
F
Nabet
P
P
D000641:Ammonia / Q000302:isolation & purification / Q000378:metabolism* / Q000633:toxicity; D000818:Animals; D001709:Biotechnology; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry / Q000633:toxicity; D003584:Cytological Techniques / Q000295:instrumentation; D005374:Filtration / Q000295:instrumentation / Q000379:methods; D018698:Glutamic Acid / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D006825:Hybridomas / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D007474:Ion Exchange; D008567:Membranes, Artificial; D051379:Mice; D017641:Zeolites
D003470:Culture Media; D008567:Membranes, Artificial; D005973:Glutamine; C083175:clinoptilolite; D017641:Zeolites; D018698:Glutamic Acid; D000641:Ammonia