Characteristics of five cell types appearing during in vitro culture of embryonic material from Drosophila melanogaster

Development ◽  
1970 ◽  
Vol 23 (1) ◽  
pp. 53-69
Author(s):  
Glen Shields ◽  
James H. Sang

Although attempts have been made over a number of decades to achieve successful tissue culture of Drosophila material, progress has been held back until recently by a lack of basic information on the chemical and physiological characteristics of the haemolymph of the organism necessary for a reasoned formulation of a culture medium. In 1963, Begg & Cruickshank published details of the mineral composition, osmotic pressure and pH of the haemolymph of third instar larvae, and this has provided the basis for a more satisfactory approach, as reflected in an increasing amount of fruitful work since reported. Much of this work has been done with embryonic material. Horikawa & Fox (1964) claimed continuous multiplication of a small type of early embryonic cell; Lesseps (1965) described re-aggregation of dissociated embryonic cells in culture, with possible development of muscle cells, nerve cells and oenocytes in the aggregates.


Blood ◽  
1974 ◽  
Vol 44 (1) ◽  
pp. 77-85 ◽  
Author(s):  
Allan J. Erslev

Abstract Normal rabbits exposed to 0.4 atmospheric pressure for 3 hr will generate about 40-60 U of erythropoietin during a subsequent 3-hr period. If the kidneys were removed from 3-hr hypoxic animals, washed carefully, and perfused for 3 hr by recirculation with a serum-tissue culture mixture, each kidney generated about 14 U of erythropoietin in vitro. Perfusion of normal kidneys did not result in the production of erythropoietin, and only small amounts were generated if the perfusate contained Puromycin. Three-hour hypoxic kidneys perfused for 3 hr with a serum-free tissue culture medium were found to generate about 8 U of erythropoietin per kidney and similar kidneys perfused with saline about 1 U. These results indicate that erythropoietin is synthesized by kidney tissue and not produced by enzymatic activation of a plasma substrate.



1988 ◽  
Vol 90 (1) ◽  
pp. 73-77
Author(s):  
A. Harris ◽  
L. Coleman

The establishment of a tissue-culture system for epithelial cells derived from human foetal pancreas has recently been reported. Further analyses have now been made on these cells in vitro, together with parallel investigation of the distribution of different cell types within the intact foetal pancreas. Results support the view that the cultured cells are ductal in origin and nature. Pancreatic epithelial cell cultures have also been established from foetuses with cystic fibrosis.



Development ◽  
1975 ◽  
Vol 34 (2) ◽  
pp. 387-405
Author(s):  
S. A. Iles ◽  
M. W. McBurney ◽  
S. R. Bramwell ◽  
Z. A. Deussen ◽  
C. F. Graham

Mouse eggs were activated with hyaluronidase in vitro and subsequently transferred to the oviduct. In the female reproductive tract they formed morulae and blastocysts which died soon after implantation. Haploid blastocysts were transferred beneath the kidney capsule and here some formed disorganized egg-cylinder structures in a week. Morulae and blastocysts from haploid and diploid parthenogenones were also transferred beneath the testis capsule. Two to four months later the growths which had formed were sectioned. They contained neural tissue, pigment, keratinized epithelium, glandular epithelium, ciliated epithelium, cartilage, bone, muscle, adipose tissue, and haemopoietic tissue. The range of cell types was similar to that produced by fertilized control blastocysts except that the parthenogenones did not form identifiable yolk-sac carcinoma or embryonal carcinomacells. The growths from haploid and diploid parthenogenones in the testis were stained with Feulgen and their DNA content measured. Growths from diploid embryos contained the normal diploid amount of DNA while growths from haploid embryos contained less than this amount. Cell cultures were prepared from the growths. The cells which were investigated contained no Y chromosome, suggesting that they were derived from the embryonic cells rather than the cells of the male host. These cells contained a near diploid chromosome number, although some of them were originally derived from haploid embryos.



1958 ◽  
Vol 36 (1) ◽  
pp. 171-184 ◽  
Author(s):  
Arthur E. Pasieka ◽  
Helen J. Morton ◽  
Joseph F. Morgan

Freshly-explanted chick embryonic kidney, chick embryonic liver, and trypsinized monkey kidney cortex cells have been cultivated in vitro in completely synthetic medium M 150. The amino acid changes in the nutrient medium during cultivation of these tissues have been studied by paper chromatography. A characteristic pattern of amino acid uptake and accumulation in the used culture medium has been demonstrated with each type of tissue culture. It has also been shown that, while the amino acid changes in the medium are different with each type of tissue culture, all cultures examined removed adenine from the medium and liberated small amounts of material thought to be hypoxanthine.



1993 ◽  
Vol 14 (9) ◽  
pp. 527-529 ◽  
Author(s):  
R. Wesley Farr ◽  
Cheryl Walton

AbstractObjective:To study the ability of a medical waste disposal process using chlorine dioxide to inactivate human immunodeficiency virus type 1 (HIV 1).Design:Stock HIV-1 (HTLV-IIIB strain) was treated with chlorine dioxide under the following settings: cell culture medium alone, culture medium with 25% blood, culture medium with medical supplies treated by the Condor machine (Winfield Environmental Corp., Escondido, CA). MT-2 cells in 96-well tissue culture plates were inoculated with serial tenfold dilutions of treated and untreated HIV-1. Cytopathic effect was read on day five, and the TCID50 (50% tissue culture infectious dose) was calculated.Results:Treatment of HIV-1 with chlorine dioxide in culture medium alone resulted in a 5.25 log10 reduction in TCID50. Treatment of HIV-1 with chlorine dioxide in the presence of 25% blood caused a 6.25 log10 reduction in HIV-1 infectivity Treatment of HIV-1 with chlorine dioxide in the presence of medical supplies treated in the Condor machine resulted in a 4.75 log10 reduction in HIV infectivity.Conclusions:Chlorine dioxide inactivated HIV-1 in vitro. Chlorine dioxide inactivated HIV-1 in the presence of blood and in the presence of medical supplies under conditions that simulated the conditions existing in the Condor machine.



2000 ◽  
Vol 74 (3) ◽  
pp. 203-208 ◽  
Author(s):  
K. Buchmann ◽  
C.V. Nielsen ◽  
J. Bresciani

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.



1952 ◽  
Vol 96 (2) ◽  
pp. 137-150 ◽  
Author(s):  
Emanuel Suter

A technique has been described for the cultivation in vitro of normal mononuclear cells on glass slides in a liquid medium. Under these conditions the monocytes transformed into macrophages which proliferated as in ordinary tissue culture. These cultures of monocytes could be infected with tubercle bacilli. The numbers of stainable tubercle bacilli within the monocytes increased steadily in cultures infected with virulent or attenuated strains. Evidence is given to support the view that this increase in numbers of bacilli was due to intracellular multiplication. There was no evidence of intracellular bacillary multiplication in cultures infected with an avirulent strain. Tubercle bacilli multiplying within phagocytes in vitro exert a damaging effect upon the host cells. The damage was most obvious in cells infected with a virulent strain. Tubercle bacilli within phagocytes were protected against the bacteriostatic effect of streptomycin added in a concentration of 5 γ per ml. of culture medium. This permitted the use of streptomycin in infected cultures to prevent extracellular multiplication of the bacilli.



2001 ◽  
Vol 280 (5) ◽  
pp. R1555-R1563 ◽  
Author(s):  
Robert M. Douglas ◽  
Tian Xu ◽  
Gabriel G. Haddad

We and others recently demonstrated that Drosophila melanogaster embryos arrest development and embryonic cells cease dividing when they are deprived of O2. To further characterize the behavior of these embryos in response to O2 deprivation and to define the O2-sensitive checkpoints in the cell cycle, embryos undergoing nuclear cycles 3–13 were subjected to O2deprivation and examined by confocal microscopy under control, hypoxic, and reoxygenation conditions. In vivo, real-time analysis of embryos carrying green fluorescent protein-kinesin demonstrated that cells arrest at two major points of the cell cycle, either at the interphase (before DNA duplication) or at metaphase, depending on the cell cycle phase at which O2 deprivation was induced. Immunoblot analysis of embryos whose cell divisions are synchronized by inducible String (cdc25 homolog) demonstrated that cyclin B was degraded during low O2 conditions in interphase-arrested embryos but not in those arrested in metaphase. Embryos resumed cell cycle activity within ∼20 min of reoxygenation, with very little apparent change in cell cycle kinetics. We conclude that there are specific points during the embryonic cell cycle that are sensitive to the O2 level in D. melanogaster. Given the fact that O2deprivation also influences the growth and development of other species, we suggest that similar hypoxia-sensitive cell cycle checkpoints may also exist in mammalian cells.





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.



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