Extra- and Intracellular Metabolite Concentrations for Murine Hybridoma Cells

Overview

Journal Appl Microbiol Biotechnol

Specialties Biomedical Engineering
Biotechnology
Microbiology

Date 1992 Feb 1

PMID 1368066

Citations 7

Authors

G Schmid

H W Blanch

Affiliations

Soon will be listed here.

Abstract

Intra- and extracellular metabolite concentrations were determined for hybridoma cells grown in tissue culture flasks in batch culture. Significant differences were found for intracellular lactate and amino acid concentrations depending on the culture medium. AB2-143.2 cells cultivated in Dulbecco's modified Eagle's (DME) medium supplemented with 50 mM lactate exhibited intracellular lactate and glutamine levels of 40 mM and 4 mM, respectively, whereas cells grown in standard DME medium had low intracellular glutamine and 20 mM lactate concentrations. For cells cultivated in medium supplemented with 6 mM pyruvate, intracellular lactate levels were estimated at approx. 40 mM, but these cells also showed an increased intracellular alanine concentration of 22 mM. The higher alanine pools probably result from increased transamination of pyruvate under these conditions.

Citing Articles

Impact of Influenza A Virus Infection on Growth and Metabolism of Suspension MDCK Cells Using a Dynamic Model.

Ramos J, Bissinger T, Genzel Y, Reichl U Metabolites. 2022; 12(3).

PMID: 35323683 PMC: 8950586. DOI: 10.3390/metabo12030239.

A mechanistic kinetic description of lactate dehydrogenase elucidating cancer diagnosis and inhibitor evaluation.

Tang P, Xu J, Oliveira C, Li Z, Liu S J Enzyme Inhib Med Chem. 2017; 32(1):564-571.

PMID: 28114833 PMC: 6010104. DOI: 10.1080/14756366.2016.1275606.

Effects of dissolved oxygen levels and the role of extra- and intracellular amino acid concentrations upon the metabolism of mammalian cell lines during batch and continuous cultures.

Heidemann R, Lutkemeyer D, Buntemeyer H, Lehmann J Cytotechnology. 2012; 26(3):185-97.

PMID: 22358616 PMC: 3449553. DOI: 10.1023/A:1007917409455.

Profiling metabolites and peptides in single cells.

Rubakhin S, Romanova E, Nemes P, Sweedler J Nat Methods. 2011; 8(4 Suppl):S20-9.

PMID: 21451513 PMC: 3312877. DOI: 10.1038/nmeth.1549.

Influence of targeted asparagine starvation on extra- and intracellular amino acid pools of cultivated Chinese hamster ovary cells.

Seewoster T, Lehmann J Appl Microbiol Biotechnol. 1995; 44(3-4):344-50.

PMID: 8597537 DOI: 10.1007/BF00169927.

References

Darmaun D, Matthews D, Desjeux J, Bier D . Glutamine and glutamate nitrogen exchangeable pools in cultured fibroblasts: a stable isotope study. J Cell Physiol. 1988; 134(1):143-8. DOI: 10.1002/jcp.1041340118. View

Wheatley D, Inglis M, Malone P . The concept of the intracellular amino acid pool and its relevance in the regulation of protein metabolism, with particular reference to mammalian cells. Curr Top Cell Regul. 1986; 28:107-82. DOI: 10.1016/b978-0-12-152828-7.50005-6. View

Reitzer L, Wice B, Kennell D . Evidence that glutamine, not sugar, is the major energy source for cultured HeLa cells. J Biol Chem. 1979; 254(8):2669-76. View

Eagle H, Piez K, Levy M . The intracellular amino acid concentrations required for protein synthesis in cultured human cells. J Biol Chem. 1961; 236:2039-42. View

Sambuy Y, Bittles A . The effects of in vitro ageing on the composition of the intracellular free amino acid pool of human diploid fibroblasts. Mech Ageing Dev. 1982; 20(4):279-87. DOI: 10.1016/0047-6374(82)90096-3. View

Myara I, Charpentier C, Gautier M, LEMONNIER A . Cell density affects prolidase and prolinase activity and intracellular amino acid levels in cultured human cells. Clin Chim Acta. 1985; 150(1):1-9. DOI: 10.1016/0009-8981(85)90305-5. View

KUCHLERJ R . THE FLEXIBLE NATURE OF THE AMINO ACID POOL IN L STRAIN FIBROBLASTS. Proc Soc Exp Biol Med. 1964; 116:20-5. View

Mohri T . Effects of cortisol and 19-nortestosterone on the free amino acid pools and amino acid uptake of cultured cells. Endocrinology. 1967; 81(3):454-60. DOI: 10.1210/endo-81-3-454. View

Butler J, Spielberg S . Decrease of intracellular cystine content in cystinotic fibroblasts by inhibitors of gamma-glutamyl transpeptidase. J Biol Chem. 1981; 256(9):4160-3. View

10.

Schmid G, Wilke C, Blanch H . Continuous hybridoma suspension cultures with and without cell retention: kinetics of growth, metabolism and product formation. J Biotechnol. 1992; 22(1-2):31-40. DOI: 10.1016/0168-1656(92)90130-2. View

11.

Goh C, Dumbroff E, Lepock J . Amino acid pools in CHL V79 cells during induction of thermotolerance: reduction in free intracellular glutamine. J Cell Physiol. 1988; 135(1):139-44. DOI: 10.1002/jcp.1041350120. View

12.

Hod Y, Hershko A . Relationship of the pool of intracellular valine to protein synthesis and degradation in cultured cells. J Biol Chem. 1976; 251(14):4458-7. View

13.

Delhotal B, Lemonnier F, Couturier M, LEMONNIER A . Influence of two cell harvesting methods on intracellular ATP and amino acid concentrations in human fibroblast cultures. Biochimie. 1983; 65(2):121-5. DOI: 10.1016/s0300-9084(83)80182-5. View

14.

Piez K, Eagle H . The free amino acid pool of cultured human cells. J Biol Chem. 1958; 231(1):533-45. View

15.

Melancon S, Tayco J, NADLER H . The free amino acid pool of cultivated human skin fibroblasts. Proc Soc Exp Biol Med. 1972; 141(1):391-5. DOI: 10.3181/00379727-141-36783. View

16.

Fernandez E, Mancuso A, Murphy M, Blanch H, Clark D . Nuclear magnetic resonance methods for observing the intracellular environment of mammalian cells. Ann N Y Acad Sci. 1990; 589:458-75. DOI: 10.1111/j.1749-6632.1990.tb24264.x. View

17.

Eagle H . Amino acid metabolism in mammalian cell cultures. Science. 1959; 130(3373):432-7. DOI: 10.1126/science.130.3373.432. View

18.

Petronini P, Tramacere M, Mazzini A, Kay J, Borghetti A . Control of protein synthesis by extracellular Na+ in cultured fibroblasts. J Cell Physiol. 1989; 140(2):202-11. DOI: 10.1002/jcp.1041400203. View

19.

Holopainen I, Oja S, Marnela K, Kontro P . Free amino acids of rat astrocytes in primary culture: changes during cell maturation. Int J Dev Neurosci. 1986; 4(5):493-6. DOI: 10.1016/0736-5748(86)90031-6. View

20.

Patel A, Hunt A . Concentration of free amino acids in primary cultures of neurones and astrocytes. J Neurochem. 1985; 44(6):1816-21. DOI: 10.1111/j.1471-4159.1985.tb07173.x. View