Home News About Us Contact Contributors Disclaimer Privacy Policy Help FAQ

Home
Search
Quick Search
Advanced
Fulltext
Browse
Collections
Persons
My eDoc
Session History
Login
Name:
Password:
Documentation
Help
Support Wiki
Direct access to
document ID:


          Institute: MPI für Dynamik komplexer technischer Systeme     Collection: Bioprocess Engineering     Display Documents



  history
ID: 221815.0, MPI für Dynamik komplexer technischer Systeme / Bioprocess Engineering
Substitution of glutamine by pyruvate to reduce ammonia formation and growth inhibition of mammalian cells
Authors:Genzel, Y.; Ritter, J. B.; König, S.; Alt, R.; Reichl, U.
Language:English
Date of Publication (YYYY-MM-DD):2005
Title of Journal:Biotechnology Progress
Volume:21
Issue / Number:1
Start Page:58
End Page:69
Review Status:Peer-review
Audience:Experts Only
Abstract / Description:In mammalian cell culture technology glutamine is required for biomass synthesis and as a major energy source together with glucose. Different pathways for glutamine metabolism are possible, resulting in different energy output and ammonia release. The accumulation of ammonia in the medium can limit cell growth and product formation. Therefore, numerous ideas to reduce ammonia concentration in cultivation broths have been developed. Here we present new aspects on the energy metabolism of mammalian cells. The replacement of glutamine (2 mM) by pyruvate (10 mM) supported cell growth without adaptation for at least 19 passages without reduction in growth rate of different adherent commercial cell lines (MDCK, BHK21, CHO-K1) in serum-containing and serum-free media. The changes in metabolism of MDCK cells due to pyruvate uptake instead of glutamine were investigated in detail (on the amino acid level) for an influenza vaccine production process in large-scale microcarrier culture. In addition, metabolite profiles from variations of this new medium formulation (1-10 mM pyruvate) were compared for MDCK cell growth in roller bottles. Even at very low levels of pyruvate (1 mM) MDCK cells grew to confluency without glutamine and accumulation of ammonia. Also glucose uptake was reduced, which resulted in lower lactate production. However, pyruvate and glutamine were both metabolized when present together. Amino acid profiles from the cell growth phase for pyruvate medium showed a reduced uptake of serine, cysteine, and methionine, an increased uptake of leucine and isoleucine and a higher release of glycine compared to glutamine medium. After virus infection completely different profiles were found for essential and nonessential amino acids.
External Publication Status:published
Document Type:Article
Communicated by:Udo Reichl
Affiliations:MPI für Dynamik komplexer technischer Systeme/Bioprocess Engineering
Identifiers:LOCALID:5/05
URL:http://dx.doi.org/10.1021/bp049827d
The scope and number of records on eDoc is subject to the collection policies defined by each institute - see "info" button in the collection browse view.