The Metabolism of Animal Tissues Cultivated in Vitro: Iii. Amino Acid Metabolism of Strain L Cells in Completely Synthetic Media

Pasieka, Arthur E.; Morton, Helen J.; Morgan, Joseph F.

doi:10.1139/o58-082

Cited by 17 publications

(4 citation statements)

References 12 publications

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“…According to our results the L strain cells in medium 199 plus 2% horse serum metabolized the amino acids somewhat differently than when maintained in a completely defined medium (Pasieka et al, 1958b). On the other hand, perhaps our results can be explained on the basis of observations of Pasieka et al that no amino acids accumulated when glutamine was omitted from the medium.…”

Section: Discussionsupporting

confidence: 54%

“…Information on the amino acid metabolism of cell cultures has been summarized recently by Morgan (1958), Eagle (1959), and Lucy (1960). With regard to the L strain of fibroblasts the cells of clone 929 take up some amino acids and accumulate others during maintenance in a defined medium (last two columns in Table 8, from Pasieka, Morton, and Morgan, 1958b). According to Sinclair (1960), this strain in a completely defined medium (Waymouth's) showed a rapid uptake of glutamine and accumulation of glutamic acid; ammonia also accumulated, even in the cell-free incubated medium.…”

Section: Discussionmentioning

confidence: 99%

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Metabolism of Animal Cells Infected With Mycoplasma

Powelson

1961

J Bacteriol

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Park, Calif.). Metabolism of animal cells infected with mycoplasma. J. Bacteriol. 82:288-297. 1961.-The effect of pleuropneumonia-like organisms (PPLO) upon the metabolism of tissue cultures was tested by comparing the assimilation and accumulation of the amino acids in the medium during growth and maintenance of monolayers of mouse fibroblasts (L strain) and human bone marrow cells (Mox). This preliminary study indicates that PPLO do alter the amino acid metabolism of animal cells. The observed changes in metabolic patterns shown by the infected fibroblast cultures did not mirror the metabolic patterns of the PPLO in the medium alone. Different strains of animal cells showed different responses to one PPLO strain, and different strains of PPLO caused different responses in one strain of animal cells. The PPLO did not grow in the tissue culture medium (no. 199 plus 2% horse serum and 20 to 40 units of penicillin/ml) nor in spent culture fluids and rapidly died off at 37 C but survived for months at 4 C. The altered metabolism of the infected tissue cultures appeared to reflect a true host-parasite interaction.

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Section: Discussionsupporting

confidence: 54%

Section: Discussionmentioning

confidence: 99%

Metabolism of Animal Cells Infected With Mycoplasma

Powelson

1961

J Bacteriol

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“…xagler. ) Laboratory of Hygiene, Department of Il'ational Health and Welfare, Ottawa, Canada. Paper chromatographic studies on the amino acid metabolism of animal tissues in vitro (1) have shown that marked changes in amino acid content of synthetic medium JI 1 50 ( 2.3 ) occur during the cultivation period.…”

Section: Specific Determination Of Hydroxy-l-proline In Biological Mamentioning

confidence: 99%

“…Full details of the chromatographic procedures have been reported previously ( 1,9,10). Chick embryo extract was prepared by grinding 11-*The cooperation of Mr. C. E. Kerr, of this Department, in making the photographic preparations, is gratefully acknowledged.…”

Section: Specific Determination Of Hydroxy-l-proline In Biological Mamentioning

confidence: 99%

Specific Determination of Hydroxy-L-Proline in Biological Materials.

Pasieka¹,

Morgan²

1956

Experimental Biology and Medicine

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Paper chromatographic studies on the amino acid metabolism of animal tissues in vitro (1) have shown that marked changes in amino acid content of synthetic medium JI 1 50 ( 2.3 ) occur during the cultivation period.Medium M 150 contains 60 ingredients, including 20 amino acids, in a modified Tyrode's solution(4) and its complexity made it difficult to detect changes in amino acids that were present in low concentrations or that did not separate completely on the chromatograms. Particular difficulty was encountered with hydroxyproline, which reacted only slightly on development with ninhydrin( S ) , isatin ( 6 ) , vanillin (7) or isatin and p-dimethylaminobenzaldehyde (8). For this reason, an effort was made to develop a selective method for hydroxyproline that could be applied in the presence of high concentrations of other amino acids. The present communication reports such a specific method, based on a modification of the conventional ninhydrin procedure. The application of this method to complex synthetic media and to deproteinized chick embryo extract is presented.Samples of media for analysis (5.0 ml) were concentrated to dryness in vacuo over H2S04, reconstituted in 0.2 ml of deionized water and 10 ii portions used without desalting. One-dimensional descending chromatograms (Schleicher and Schuell S o . 597 or Whatman No. 1 paper) were developed for 2 successive 18-hour periods in either n-butanol-acetic acid-water or n-butanol-ethanol-water.The chromatograms were dried at 110°C for 2 to 3 minutes and sprayed with 0.4% ninhydrin in water-saturated n-butanol or 95% ethanol. Full details of the chromatographic procedures have been reported previously ( 1,9,10). Chick embryo extract was prepared by grinding 11-*The cooperation of Mr. C. E. Kerr, of this Department, in making the photographic preparations, is gratefully acknowledged. Methods.~-_-day-old embryos and centrifuging off the tissue pulp( 11). The supernatant extract was adjusted to pH 4.0 with 1 r\' H2S04, precipitated proteins removed by centrifuging, and the pH readjusted to 7.2. Two volumes of acetone were added, the mixture allowed to stand overnight in the refrigerator, and centrifuged. The final supernatant was concentrated to dryness in the usual manner and used for analysis. Synthetic medium M 150 was prepared as described previously( 2,3). All chemicals and reagents were of the highest purity obtainable. Spectrophotometric measurements were made by cutting appropriate areas from the developed chromatograms and fixing them to the inner walls of 1 cm Corex cells. Determinations were then made in a Beckman, Model DC. Ultraviolet examinations were carried out with a laboratory hand lamp (short wave model, 2537Results. Development of a specific color with hydroxy-L-proline and ninhydrin, Previous studies on the determination of phenylalanine in synthetic medium 31 lSO(10) showed that dipping ninhydrin-developed chromatograms in 1 % sodium bicarbonate produced a stable blue color characteristic of this amino acid. At the same time, this alkali tre...

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The metabolism of L‐glutamate and L‐5‐Carboxypyrrolidone by mouse cells (NCTC clone 929) under conditions of defined nutrition

Kitos¹,

Waymouth²

1966

Journal Cellular Physiology

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The utilization of L-glutamate by clone 929 mouse cells growing in a synthetic medium, MAL 294/2, was studied with the aid of carbon-14 labeled L-glutamate. The rate of consumption of extracellular glutamate was rapid even though the extracellular concentration of this substance has been found to remain constant or to increase. The rate of uptake during an interval of otpimal growth was calculated to be approximately 42 mpmoles/mg of cell protein per hour.Among the metabolic products that are derived from the carbon of glutamate and secreted from the cells are carbon dioxide, lactic acid, proline, alanine, alpha-ketoglutaric acid and 5-carboxypyrrolidone. Aspartic acid, although produced by the cells in amounts sufficient to meet the needs for growth, does not appear as an extracellular product of glutamate metabolism. Extracts of L cells were found to exhibit four times as much glutamic-oxaloacetic as glutamic-pyruvic transaminase activities. Failure to secrete aspartic acid must not be due to a deficiency in the transaminase. The transaminase concentrations are apparently not affected by variations in the concentrations of aspartic acid and alanine in the medium, both of which are absent from MAL 294/2. 5-Carboxypyrrolidone, although produced from L-glutamate by L cells, is metabolically inert in this system. Likewise, mouse fetal lung cells, cultured in a similar way, use glutamic acid as extensively as L cells and fail to metabolize exogenous 5-carboxypyrrolidone.

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The Metabolism of Animal Tissues Cultivated in Vitro: Iii. Amino Acid Metabolism of Strain L Cells in Completely Synthetic Media

Cited by 17 publications

References 12 publications

Metabolism of Animal Cells Infected With Mycoplasma

Metabolism of Animal Cells Infected With Mycoplasma

Specific Determination of Hydroxy-L-Proline in Biological Materials.

The metabolism of L‐glutamate and L‐5‐Carboxypyrrolidone by mouse cells (NCTC clone 929) under conditions of defined nutrition

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