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

Healy, George M.; Fisher, Dorothy C.; Parker, Raymond C.

doi:10.1139/o54-035

Cited by 48 publications

(4 citation statements)

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“…Good growth in vitro is a function of various factors (28). Although the synthetic media used are very suitable for enzyme work because they do not contain enzymes (7-9) the growth of tissues is not so great with the majority of these mixtures (11,50). Suspended kidney fragments with complete and incomplete media, and human amniofic cells grown in complex natural nutrients are under study, to correlate the findings on different materials (13).…”

Section: Discussionmentioning

confidence: 99%

Comparative Biochemical Studies on Normal and on Poliomyelitis Virus-Infected Tissue Cultures

Kovács

1956

The Journal of Experimental Medicine

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A survey of the literature of tissue cultivation does not disclose much information on enzymes in this particular branch of experimental biology (1). The majority of writers dealing with phosphomonoesterases in cultivated cells (2, 3), with the exception of Fell and Robinson (4, 5), applied histochemical tests (6), which do not permit quantitative evaluation. We followed the activities of various enzyme systems in normal tissue cultures (TC) by biochemical methods (7,8). Using similar techniques striking changes were observed in nine enzyme systems of cells infected with various strains of poliomyelitis virus and the findings published in a preliminary report (9). The present publication deals exclusively with various aspects of phosphatase activities in virus-inoculated TC and in normal controls. Previous works in this field were made mostly on organ homogenates of experimental animals (10), containing all the products of inflammatory reactions of the host. Our contribution on relatively simple systems concerns the mechanism of drastic modifications in cell physiology during poliomyelitis infection, in vitro. MethodsOnly the variations in the handling of TC will be mentioned here; for more information and the general methods the reader is referred to the previous papers of this series (7-9).Explants.--Tissue cultivation, virus production, and titration were made by the Poliomyelitis Assay Department. I Most of the experiments were carried out on roller tubes of trypsin~zed rhesus monkey kidney cortex, grown about 14 days with synthetic nutrient 199 (11) and later with a modified form of it, termed medium 597 (7). The roller tube cultures were inoculated with various dilutions of virus, generally 24 hours after the second fluid change, and were incubated in roller drums at 37°C. for 6 days (12). For calculation of the titre K~.rber's method was used (13) and virus concentration was expressed in negative logarithms (14). Specially designed experiments were run with large inocula

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Section: Discussionmentioning

confidence: 99%

Comparative Biochemical Studies on Normal and on Poliomyelitis Virus-Infected Tissue Cultures

Kovács

1956

The Journal of Experimental Medicine

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“…The following strains of cells were propagated serially as described previously (7-9) : human fibroblasts, strains FS4-705 and U12-705 (8), rabbit fibroblasts, strains RM3-56, RS1-56, and RT6-56 (9) and mouse fibroblasts, strain L-705 (10). Medium 705 is composed of solution 703 (11) supplemented with 5 per cent (V/V) chick embryo extract (CEE, 7) and 20 per cent normal horse serum (NHS) and medium 56 contains 5 per cent CEE, 10 per cent NHS, and 85 per cent solution S18 (12). Dialyzed chick embryo extract (DCEE) and dialyzed horse serum (DHS) are prepared by dialysis (12).…”

Section: Methodsmentioning

confidence: 99%

The Role of Carbon Dioxide as an Essential Nutrient for Six Permanent Strains of Fibroblasts

Swim

Parker

1958

The Journal of Cell Biology

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The results of these studies demonstrate that carbon dioxide is required for the growth and maintenance of strains of fibroblasts derived from human tissues, strains FS4-705 and U12-705, from mouse tissue, strain L-705, and from rabbit tissues, strains RM3-56, RS1-56, and RT-56 in a chemically defined medium containing phosphite buffer in place of bicarbonate and supplemented with dialyzed serum and dialyzed embryo extract. Under these conditions, the cells fail to proliferate at a significant rate and begin to degenerate within 5 to 10 days when the flasks are not stoppered. Suflficient carbon dioxide is produced by the ceils to promote growth as indicated by the fact that maximal proliferation is obtained in the same phosphite media when stoppered flasks are employed. With the exception of RS1-56, all the remaining strains tested can be propagated serially in open flasks containing phosphite medium prepared with whole serum and embryo extract. The rate of growth under these conditions, however, is only one-half to onethird that obtained in stoppered flasks containing phosphite medium or the conventional bicarbonate medium.An earlier report from this laboratory described the propagation of several strains of mammalian cells in media containing fixed buffers such as tris(hydroxymethyl)aminomethane and glycylglycine in place of the conventional carbon dioxidebicarbonate system (1). Studies on the practical application of fixed buffers in tissue culture media were subsequently extended to include other compounds which are compatible with calcium and other cations and which are effective buffers in the p H range 7.0-8.0. In the course of these investigations, it was observed that several strains of fibroblasts proliferate at a reduced rate in certain media containing sodium phosphite (2) in place of bicarbonate when the flasks were dosed only by means of loosely fitting metal caps instead of stoppers. These observations suggested that carbon dioxide might be an essential nutrient for fibroblasts propagated in vitro. Some support for this * Aided by a grant (No. E-1547) from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, United States Public Health Service. speculation was indicated by other investigations which demonstrated that carbon dioxide is beneficial for the outgrowth of cells from explanted rat kidney (3, 4) and essential for explants of chick embryo tissue (5, 6). The results to be presented demonstrate that carbon dioxide is essential for the proliferation of six permanent strains of fibroblasts, derived from human, mouse, and rabbit tissues, when propagated in media containing dialyzed chick embryo extract and dialyzed horse serum. Materials and MethodsThe following strains of cells were propagated serially as described previously (7-9) : human fibroblasts, strains FS4-705 and U12-705 (8), rabbit fibroblasts, strains RM3-56, RS1-56, and RT6-56 (9) and mouse fibroblasts, strain L-705 (10). Medium 705 is composed of solution 703 (11) supplemented with 5 pe...

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“…The problem of the toxicity of GSH and cysteine to the lens was also considered. Healy, Fisher, and Parker (1954) (Johnson, 1936 The medium is also stable over a three-day period in regard to pH change.…”

Section: Introductionmentioning

confidence: 99%

Development of a Synthetic Medium for Rabbit Lens Culture in a Perfusion System

Schwartz¹

1960

AMA Arch Ophthalmol

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

Cited by 48 publications

References 1 publication

Comparative Biochemical Studies on Normal and on Poliomyelitis Virus-Infected Tissue Cultures

Comparative Biochemical Studies on Normal and on Poliomyelitis Virus-Infected Tissue Cultures

The Role of Carbon Dioxide as an Essential Nutrient for Six Permanent Strains of Fibroblasts

Development of a Synthetic Medium for Rabbit Lens Culture in a Perfusion System

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