Bilans nutritifs glucidique et azoté de tissus de tubercules de topinambour cultivés in vitro après irradiation gamma

1982 ◽  
Vol 60 (11) ◽  
pp. 2215-2218
Author(s):  
Janine Schaeverbeke-Sacré ◽  
Béatrice Matheron

Carbohydrate and nitrogen budgets were studied in Jerusalem artichoke tuber explants cultured in vitro after gamma irradiation (0 to 106 rads (1 rad = 10−2 J/kg)). A certain level of reducing sugars is reached and retained in all the explants and, in all cases, an increase of protein nitrogen is observed. The highly irradiated tissues have a very disturbed metabolism that involves in particular, amino acid leaching into the culture medium.

1983 ◽  
Vol 61 (5) ◽  
pp. 1448-1455 ◽  
Author(s):  
Janine Schaeverbeke-Sacré ◽  
Béatrice Matheron

DNA and RNA contents are studied in Jerusalem artichoke tuber explants cultured in vitro after gamma irradiation (0–5 × 105 rads (1 rad = 10 mGy)). The lower part of the explants is stimulated as soon as in contact with the medium. This stimulated area is still able to synthesize DNA and RNA up to 104 rads. An histological and cytological study shows that tissue neoformations can be observed up to 6000 rads in this outer area and that gamma rays seem to keep the cells in a "premitotic" state for a longer or shorter period according to the applied radiation dose.


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.


1998 ◽  
Vol 10 (3) ◽  
pp. 279 ◽  
Author(s):  
Y. G. Jung ◽  
T. Sakata ◽  
E. S. Lee ◽  
Y. Fukui

The uptake and synthesis of 19 amino acids by fresh or frozen–thawed bovine blastocysts produced by parthenogenesis (PT) or in vitro fertilization (IVF) were compared in the present study. Fresh blastocysts, 180 h after IVF or PT activation, and frozen–thawed blastocysts, 168 h old and cultured for 12 h post-thawing, were cultured in synthetic oviduct fluid medium (SOFM) containing polyvinyl alcohol (PVA) with both essential and non-essential amino acids (EAA and NEAA, respectively) (Medium 1: M1) or SOFM containing PVA with only EAA (Medium 2: M2). In Experiment 1, when fresh or frozen–thawed PT blastocysts were cultured in M1, the uptake of glutamate (in fresh only), aspartate and arginine, and the synthesis of glutamine and alanine were significantly enhanced. In the culture with M2, serine, asparagine, glutamate, glutamine, glycine, arginine and alanine were significantly taken up. It was found that the glutamine concentrations was significantly higher (P < 0.001) in the culture medium drops containing embryos than in the drops without embryos. In Experiment 2, when PT blastocysts were cultured in M1, the uptake of aspartate and synthesis of alanine were greater (P < 0.01) than those by IVF blastocysts. When M2 was used, a significant (P < 0.01) production of serine, asparagine, glutamate, glutamine and alanine, and the uptake of arginine by PT blastocysts were observed. In Experiment 3, when IVF blastocysts were cultured in M1, fresh blastocysts depleted more aspartate and glutamate, and produced more glutamine and alanine than frozen–thawed blastocysts. When cultured in M2, frozen–thawed blastocysts depleted more threonine (P < 0.01) than fresh blastocysts. These results indicate that the uptake and synthesis of amino acids were different in fresh or frozen–thawed bovine blastocysts derived from PT or IVF. These differences in amino acid metabolism may be related to the viability of the blastocysts.


1996 ◽  
Vol 8 (5) ◽  
pp. 835 ◽  
Author(s):  
T Pinyopummintr ◽  
BD Bavister

Effects of amino acids on early bovine embryo development in vitro were examined using a chemically-defined, protein-free culture medium. Bovine embryos produced in vitro were cultured from 18 h to 72 h post insemination in a simple medium containing lactate as the only energy source except for the amino acid treatments. Subsequently, embryos were transferred to TCM-199 supplemented with serum for blastocyst development to substantiate their developmental competence. Treatments were: (1) non-essential amino acids from TCM-199 (NEA); (2) essential amino acids from TCM-199 (EA); (3) NEA+EA; (4) Eagle's minimum essential medium amino acids (MEM AA); (5) 11 amino acids present in HECM-6 (11 AA); and (6) 0.2 mM glutamine (GLN). A higher proportion of embryos (percentage of inseminated ova) cleaved to the > or = 8-cell stage by 72 h post insemination in NEA (56.7%), EA (41.2%), 11 AA (40.3%) and GLN (51.1%) than in either NEA+EA (30.0%) or MEM AA (33.1%). However, after transfer to complex medium, embryos that had developed in EA, as well as those in MEM AA or NEA+EA, produced significantly fewer blastocysts (37.1%, 34.4% and 25.6% respectively) than those in NEA (56.7%), GLN (48.9%) or 11 AA (37.7%). The ability of blastocysts to hatch from their zonae pellucidae was also affected by amino acid treatment during cleavage stages. The present study indicated that the addition of NEA or GLN or 11 AA to a chemically-defined culture medium during the cleavage phase of bovine embryo development increases their subsequent ability to reach the blastocyst stage. These data have implications for understanding the nutritional needs of bovine embryos produced in vitro and for optimizing the composition of culture media to support their development.


1978 ◽  
Vol 56 (3) ◽  
pp. 167-173
Author(s):  
Joachim Sparkuhl ◽  
George Setterfield

Both nongrowing (water-incubated) and growing (hormonally stimulated) Jerusalem artichoke tuber cells contain membrane-bound (mb) ribosomes. Using a rapid flotation procedure, a membrane fraction was prepared from both types of cells. This fraction was enriched in mb ribosomes, contained NADH cytochrome c reductase activity, had RNA:phospholipid and RNA:protein ratios similar to those reported for rough microsomes from animal tissues, and supported synthesis of preinitiated proteins in vitro. Using puromycin and detergent release, vectorial transport of labelled polypeptides was measured in the in vitro system. Of proteins made by mb ribosomes from nongrowing cells, only 12% remained associated with microsome membranes following chain termination. The comparable figure for proteins from mb ribosomes of growing tissue was 42%. The membrane-associated proteins were preferentially protected from protease digestion. Some possible reasons are suggested for the correlation between cell growth and the association of newly synthesized proteins with microsomes. The role of proteins synthesized by mb ribosomes but not vectorially transported, in both growing and nongrowing cells, is unknown.


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.


1958 ◽  
Vol 36 (1) ◽  
pp. 771-782 ◽  
Author(s):  
Arthur E. Pasieka ◽  
Helen J. Morton ◽  
Joseph F. Morgan

Strain L cells, of mouse fibroblastic origin, have been cultivated in vitro in completely synthetic medium M 150 and in various modifications of this medium. The amino acid changes in the nutrient medium during cell cultivation have been studied by paper chromatography. A characteristic pattern of amino acid uptake and accumulation in the medium has been found. No change in the alanine concentration was observed but omission of alanine from the culture medium resulted in its accumulation in appreciable amounts. Omission of glutamic acid did not alter the pattern of amino acid changes by the cells. Omission of glutamine increased the uptake of amino acids and prevented amino acid accumulation. Omission of both glutamic acid and glutamine resulted in a virtual cessation of amino acid changes in the culture medium. Strain L cells decreased the adenine content of the medium and produced small amounts of hypoxanthine. These changes were not affected by alterations in the amino acid content of the medium. Omission of glutamic acid and glutamine from the culture medium did not cause an appreciable decrease in cell population or apparent degeneration of the cultures over a 30-day period.


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