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: 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: 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.

Studies on the Mechanism of Substrate Induction and L-Cyst(e)ine Repression of Alkaline Phosphatase in Mammalian Cell Cultures

1967 ◽  
Vol 2 (4) ◽  
pp. 545-555
Author(s):  
M. J. GRIFFIN ◽  
R. P. COX
Keyword(s):  
Tissue Culture ◽  
Alkaline Phosphatase ◽  
Cell Cultures ◽  
Phosphate Esters ◽  
Kidney Cell Line ◽  
Monkey Kidney Cell ◽  
African Green Monkey Kidney ◽  
Mammalian Cell Cultures ◽  
Green Monkey ◽  
Substrate Induction

The mechanisms of substrate induction and L-cyst(e)ine repression of alkaline phosphatase were studied in tissue culture using an established African green monkey kidney cell line (BS-C-I). L-Cyst(e)ine repression and substrate induction are mutually antagonistic. Evidence is presented which suggests that the increase in alkaline phosphatase levels induced by mono-phosphate esters may in part be due to protection of the enzyme from cellular degradation, while L-cyst(e)ine is believed to act either by repressing the synthesis of the enzyme or by selectively increasing its catabolism.

A second rubella vaccine

1971 ◽  
Vol 9 (13) ◽  
pp. 52-52
Keyword(s):  
Tissue Culture ◽  
Pregnant Women ◽  
Rabbit Kidney ◽  
Kidney Cells ◽  
Animal Cells ◽  
Rubella Vaccine ◽  
Human Diploid Cells ◽  
Diploid Cells

Available vaccines used in protection against rubella employ strains of virus attenuated by passage in animal cells. The first vaccine that was introduced in Britain, the Cendehill vaccine (Cendevax),1 is grown on tissue culture of rabbit kidney cells. Recently another rubella vaccine (Almevax live attenuated, Wistar RA 27/3), grown on human diploid cells, has been introduced. This vaccine induces antibodies as effectively as earlier vaccines, and the indications for its use are similar to those for the Cendehill vaccine.1 2 It is very important that pregnant women, women who might be pregnant, or women likely to become pregnant within 2 months, must not be vaccinated.

In vitro interactions between epithelial cells and Gyrodactylus derjavini

2000 ◽  
Vol 74 (3) ◽  
pp. 203-208 ◽  
Author(s):  
K. Buchmann ◽  
C.V. Nielsen ◽  
J. Bresciani
Keyword(s):  
Tissue Culture ◽  
Epithelial Cells ◽  
Cell Cultures ◽  
Culture Medium ◽  
Epithelioma Papulosum Cyprini ◽  
Skin Responses ◽  
Enzyme Reactivity ◽  
In Vitro Interactions

AbstractSkin responses of fish to various parasites have been shown to involve various immunologically competent cells producing factors which guide the reactions of epithelial cells. However, the present study has demonstrated that a monoculture of epithelial cells has the ability to encapsulate and partially degrade ectoparasites without involvement of leukocytes. The ectoparasitic monogeneanGyrodactylus derjavini was kept on a monolayer of Epithelioma Papulosum Cyprini (EPC) cells in 24-well multidishes supplied with tissue culture medium. Gyrodactylus derjavini did not reproduce but survived an incubation period of up to139 h in the system. Due to sterile conditions, dead gyrodactylids were not subjected to microbial degradation and remained intact for several weeks. However, at 40 days G. derjavini was overgrown by EPC-cells and became partly degraded during the following 15 days. Analysis of enzyme reactivity in EPC-cells showed reactions for ten enzymes including esterases, amidases, phosphatases and phosphohydrolases. No marked differences for the ten enzymes between cell cultures with and without the ectoparasites were found but it cannot be excluded that some of these enzymes took part in parasite degradation. The study showed the in vitro capability of epithelial cells to interact, encapsulate and degrade G. derjavini without the involvement of leukocytes. This response probably is non-specific and will not exclude that various immunocompetent cells and their products normally optimize and accelerate elimination of invading parasites in vivo.

Human brain in tissue culture. I. Acquisition, initial processing, and establishment of brain cell cultures

1975 ◽  
Vol 161 (3) ◽  
pp. 295-306 ◽  
Author(s):  
Donald H. Gilden ◽  
Mary Devlin ◽  
Zofia Wroblewska ◽  
Harvey Friedman ◽  
Lucy Balian Rorke ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Human Brain ◽  
Cell Cultures ◽  
Brain Cell ◽  
Brain Cell Cultures ◽  
Initial Processing

Neopterin measured in serum and tissue culture supernates by a competitive enzyme-linked immunosorbant assay.

1989 ◽  
Vol 35 (7) ◽  
pp. 1467-1471 ◽  
Author(s):  
M Barak ◽  
D Merzbach ◽  
N Gruener
Keyword(s):  
Tissue Culture ◽  
Correlation Coefficient ◽  
Cell Cultures ◽  
Solid Phase ◽  
Activated Macrophages ◽  
Immunosorbant Assay ◽  
Analytical Recovery ◽  
Enzyme Linked Immunosorbant Assay

Abstract In this solid-phase competitive enzyme-linked immunosorbant assay for neopterin (a product of activated macrophages) in serum or supernates of cell cultures, we incubate antiserum to neopterin with standards or samples in the presence of solid-phase-bound conjugate of carrier-neopterin. Incubation with second antibody labeled with peroxidase then follows, before reaction with substrate. Analytical recovery, precision, and sensitivity of this method are similar to those of other immunoassays. The assay range is between 0.4 and 100 micrograms of neopterin per liter. Results obtained by this method compared with those from a conventional radioimmunoassay gave a correlation coefficient of 0.974.

Demethylation of genes in animal cells

1990 ◽  
Vol 326 (1235) ◽  
pp. 241-251 ◽  
Keyword(s):  
Tissue Culture ◽  
Germ Line ◽  
Animal Cell ◽  
Gene Activation ◽  
Housekeeping Gene ◽  
Cell Types ◽  
Embryonic Cells ◽  
Animal Cells ◽  
Developmentally Regulated ◽  
I Gene

Tissue-specific animal cell genes are usually fully methylated in the germ line and become demethylated in those cell types in which they are expressed. To investigate this process, we inserted a methylated IgG K gene into fibroblasts and lymphocytes at various stages of development. The results show that this gene undergoes demethylation only in the mature lymphocytes and therefore suggest that the ability to demethylate a gene is developmentally regulated. These studies were supported by similar experiments using the rat Insulin I gene, and in this case it appears that the cis -acting elements that control demethylation may be different from those responsible for gene activation. The ability to demethylate the housekeeping gene APRT is also under developmental control, because this occurs only in embryonic cells, both in tissue culture and in transgenic mice.

Sign in / Sign up

Export Citation Format

Share Document