NUTRITION OF ANIMAL CELLS IN TISSUE CULTURE: IX. SYNTHETIC MEDIUM No. 703

1954 ◽  
Vol 32 (3) ◽  
pp. 327-337 ◽  
Author(s):  
George M. Healy ◽  
Dorothy C. Fisher ◽  
Raymond C. Parker
Keyword(s):  
Tissue Culture ◽  
Ascorbic Acid ◽  
Chick Embryo ◽  
Synthetic Medium ◽  
Average Period ◽  
Animal Cells ◽  
Cell Life ◽  
Mouse Cells ◽  
Synthetic Media ◽  
Purines And Pyrimidines

Previous reports from this laboratory described a synthetic medium, solution 199, that contained 60 ingredients and supported cell life for an average period of four to five weeks. The assays were carried out in roller tubes in which chick embryo mesenchyme tissues were cultivated directly on the glass. More recently, the synthetic media have been improved very considerably, and Earle's replicate culture assay procedures have been adapted for use in testing them. By means of these techniques, it has been possible to make accurate quantitative experiments with new synthetic solutions some of which are capable of yielding, in one week, five- and sixfold increases in the population of replicate cultures prepared from washed suspensions of Earle's L strain mouse cells. This was acommplished by omitting the purines and pyrimidines from solution 199, by adding greatly increased levels of cysteine, ascorbic acid, and glutathione, and by the addition of certain purified coenzymes. In solution 703, which is described in detail, cultures of L strain cells have been maintained in a healthy, active state for over five months, with renewal of the medium twice a week.

NUTRITION OF ANIMAL CELLS IN TISSUE CULTURE: IX. SYNTHETIC MEDIUM No. 703

1954 ◽  
Vol 32 (1) ◽  
pp. 327-337 ◽  
Author(s):  
George M. Healy ◽  
Dorothy C. Fisher ◽  
Raymond C. Parker
Keyword(s):  
Tissue Culture ◽  
Ascorbic Acid ◽  
Chick Embryo ◽  
Synthetic Medium ◽  
Average Period ◽  
Animal Cells ◽  
Cell Life ◽  
Mouse Cells ◽  
Synthetic Media ◽  
Purines And Pyrimidines

Previous reports from this laboratory described a synthetic medium, solution 199, that contained 60 ingredients and supported cell life for an average period of four to five weeks. The assays were carried out in roller tubes in which chick embryo mesenchyme tissues were cultivated directly on the glass. More recently, the synthetic media have been improved very considerably, and Earle's replicate culture assay procedures have been adapted for use in testing them. By means of these techniques, it has been possible to make accurate quantitative experiments with new synthetic solutions some of which are capable of yielding, in one week, five- and sixfold increases in the population of replicate cultures prepared from washed suspensions of Earle's L strain mouse cells. This was acommplished by omitting the purines and pyrimidines from solution 199, by adding greatly increased levels of cysteine, ascorbic acid, and glutathione, and by the addition of certain purified coenzymes. In solution 703, which is described in detail, cultures of L strain cells have been maintained in a healthy, active state for over five months, with renewal of the medium twice a week.

NUTRITION OF ANIMAL CELLS IN TISSUE CULTURE: VII. USE OF REPLICATE CELL CULTURES IN THE EVALUATION OF SYNTHETIC MEDIA

1954 ◽  
Vol 32 (1) ◽  
pp. 306-318
Author(s):  
Raymond C. Parker ◽  
George M. Healy ◽  
Dorothy C. Fisher
Keyword(s):  
Tissue Culture ◽  
Cell Cultures ◽  
Subsequent Treatment ◽  
Relative Toxicity ◽  
Animal Cells ◽  
Washed Cell ◽  
Mouse Cells ◽  
Synthetic Media ◽  
Simple Screening ◽  
Natural Media

The replicate culture assay procedures of Earle and his associates have been adapted for use in evaluating the effectiveness of synthetic media. For this purpose, use has also been made of Earle's L strain mouse cells. Washed and continuously stirred suspensions of these or similar strains of cells may be dispensed, with reasonable assurance of uniformity, into a series of replicate cultures, the number depending on the volume of the suspension and the capacity and effectiveness of the stirring and dispensing unit. For use with synthetic media, the original procedures for the preparation and care of the replicate cultures and for their subsequent treatment for the counting of isolated, stained nuclei have been modified considerably. This paper describes the procedures that were finally adopted and also describes a relatively simple screening procedure in which washed cell suspensions may be used to advantage in making preliminary assays of synthetic media and in testing the relative toxicity or growth stimulating effects of substances added to, or derived from, natural media.

NUTRITION OF ANIMAL CELLS IN TISSUE CULTURE: VII. USE OF REPLICATE CELL CULTURES IN THE EVALUATION OF SYNTHETIC MEDIA

1954 ◽  
Vol 32 (3) ◽  
pp. 306-318 ◽  
Author(s):  
Raymond C. Parker ◽  
George M. Healy ◽  
Dorothy C. Fisher
Keyword(s):  
Tissue Culture ◽  
Cell Cultures ◽  
Subsequent Treatment ◽  
Relative Toxicity ◽  
Animal Cells ◽  
Washed Cell ◽  
Mouse Cells ◽  
Synthetic Media ◽  
Simple Screening ◽  
Natural Media

The replicate culture assay procedures of Earle and his associates have been adapted for use in evaluating the effectiveness of synthetic media. For this purpose, use has also been made of Earle's L strain mouse cells. Washed and continuously stirred suspensions of these or similar strains of cells may be dispensed, with reasonable assurance of uniformity, into a series of replicate cultures, the number depending on the volume of the suspension and the capacity and effectiveness of the stirring and dispensing unit. For use with synthetic media, the original procedures for the preparation and care of the replicate cultures and for their subsequent treatment for the counting of isolated, stained nuclei have been modified considerably. This paper describes the procedures that were finally adopted and also describes a relatively simple screening procedure in which washed cell suspensions may be used to advantage in making preliminary assays of synthetic media and in testing the relative toxicity or growth stimulating effects of substances added to, or derived from, natural media.

Nutrition of Animal Cells in Tissue Culture. X. Synthetic Medium No. 858,

1955 ◽  
Vol 89 (1) ◽  
pp. 71-77 ◽  
Author(s):  
G. M. Healy ◽  
D. C. Fisher ◽  
R. C. Parker
Keyword(s):  
Tissue Culture ◽  
Synthetic Medium ◽  
Animal Cells

Glutamine metabolism by animal cells growing in a synthetic medium

1962 ◽  
Vol 27 (2) ◽  
pp. 307-316 ◽  
Author(s):  
P.A. Kitos ◽  
R. Sinclair ◽  
C. Waymouth
Keyword(s):  
Synthetic Medium ◽  
Glutamine Metabolism ◽  
Animal Cells

The effects of D- and L-threo-chloramphenicol on the early development of the chick embryo

Development ◽  
1965 ◽  
Vol 13 (3) ◽  
pp. 341-356
Author(s):  
F. S. Billett ◽  
Rosalba Collini ◽  
Louie Hamilton
Keyword(s):  
Protein Synthesis ◽  
Amino Acid ◽  
Chick Embryo ◽  
Messenger Rna ◽  
Mammalian Cells ◽  
Animal Cells ◽  
Inhibit Protein Synthesis ◽  
Acid Complex ◽  
Amino Acid Complex ◽  
Bacterial Systems

In many bacterial systems chloramphenicol has been shown to inhibit protein synthesis (Hahn & Wisseman, 1951; Gale & Folkes, 1953). The precise mechanism of this inhibition is not clear, although the evidence suggests that the interaction of the soluble RNA-amino acid complex with the ribosomes is prevented because the attachment of the messenger RNA to the ribosomes is itself impaired (Lacks & Gros, 1959; Nathans & Lipman, 1961; Jardetsky & Julian, 1964; Julian & Jardetsky, 1964). In contrast to its effect on bacterial systems, chloramphenicol has been reported to have little or no action on the protein synthesis by cell-free extracts of mammalian cells (Rendi, 1959; Ehrenstein & Lipmann, 1961). A basis for this resistance has been proposed by Vazquez (1964), who finds that whereas bacterial ribosomes bind chloramphenicol, ribosomes from other organisms do not. Nevertheless, it cannot be stated with any confidence that chloramphenicol has no effect on the protein synthesis of animal cells.

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