Improved techniques for the induction of mammalian cell hybridization by polyethylene glycol

1976 ◽  
Vol 2 (2) ◽  
pp. 165-176 ◽  
Author(s):  
Richard L. Davidson ◽  
Park S. Gerald
1976 ◽  
Vol 2 (3) ◽  
pp. 271-280 ◽  
Author(s):  
Richard L. Davidson ◽  
Kathleen A. O'Malley ◽  
Thomas B. Wheeler

1979 ◽  
Vol 5 (2) ◽  
pp. 263-269 ◽  
Author(s):  
Sue Schneiderman ◽  
John L. Farber ◽  
Renato Baserga

1985 ◽  
Vol 78 (1) ◽  
pp. 263-271
Author(s):  
Y. Matsuya ◽  
I. Yamane

A great increase in hybridization frequency of cultured rodent cells was obtained when conventional cell fusion using 50% polyethylene glycol (PEG) was combined with a cell agglutination produced by plant lectins. The rate of appearance of hybrid colonies was found to be correlated with the extent of cell agglutination by lectin, as well as with cell fusion induced by subsequent PEG treatment. Phytohemagglutinin (PHA), wheat germ agglutinin, Wistaria floribunda agglutinin and concanavalin A were all active; the most effective was PHA. When parental cells in a monolayer were treated with PHA followed by PEG, the resulting hybridization frequency was very low because of markedly decreased viability, whereas the same cells in suspension yielded hybrid colonies at a higher rate. These results suggest that the enhancement of hybridization by PHA/PEG treatment was brought about by the ability of lectin to agglutinate cells.


1981 ◽  
Vol 36 (7-8) ◽  
pp. 593-596 ◽  
Author(s):  
Hermann Krähling

Abstract Investigations on the dose response of cell fusion, induced by ionfree aqueous polyethylene glycol (PEG) solutions, reveal distinct lowest fusogenic PEG concentrations for different permanently growing mammalian cell lines. Part of the requisite PEG can be replaced by carbo­ hydrates, preserving the fusogenity of the solutions. This discriminates two effects of PEG solutions causing cell fusion: a) cell shrinkage, the required hyperosmolality of the solutions may be provided by PEG or by carbohydrates, is supposed to cause intracellular processes necessary for consolidating polycaryons; b) membrane alterations, which can not be induced by carbo­ hydrates, enable intimate cell-cell contact via particle-free membrane areas. Depending on cell line salts can not only raise the osmolality of PEG solutions but are able to co-operate with PEG in generating membrane alterations.


1985 ◽  
Vol 78 (1) ◽  
pp. 273-282
Author(s):  
Y. Matsuya ◽  
I. Yamane

An efficient technique for hybridization of mammalian cells was developed by combining agglutination by pretreatment with polycations, such as polyarginine, and conventional polyethylene glycol(PEG)-mediated cell fusion. Polyarginine and subsequent PEG treatment resulted in markedly decreased viability in the treated cells, but addition of polyvinyl pyrrolidone or glycerol to the polyarginine prevented this cytotoxicity. Polyarginine was much more effective than polylysine or polyornithine in inducing hybridization. Other polycations, including polybrene and protamine but not DEAE-dextran, were also active in inducing hybridization. The condition of the cells at the time of polycation treatment was an important factor in the enhancement of hybridization. The condition of the cells at the time of polycation treatment was an important factor in the enhancement of hybridization. The enhancement of hybridization of cells in monolayer incubated for 2 h was much higher than that of cells incubated for 24 h. These findings suggest that polycations do not necessarily operate by agglutinating cells. The mechanism of polycation-enhanced cell hybridization is discussed.


Author(s):  
Kuixiong Gao ◽  
Randal E. Morris ◽  
Bruce F. Giffin ◽  
Robert R. Cardell

Several enzymes are involved in the regulation of anabolic and catabolic pathways of carbohydrate metabolism in liver parenchymal cells. The lobular distribution of glycogen synthase (GS), phosphoenolpyruvate carboxykinase (PEPCK) and glycogen phosphorylase (GP) was studied by immunocytochemistry using cryosections of normal fed and fasted rat liver. Since sections of tissue embedded in polyethylene glycol (PEG) show good morphological preservation and increased detectability for immunocytochemical localization of antigenic sites, and semithin sections of Visio-Bond (VB) embedded tissue provide higher resolution of cellular structure, we applied these techniques and immunogold-silver stain (IGSS) for a more accurate localization of hepatic carbohydrate metabolic enzymes.


Author(s):  
Dai Dalin ◽  
Guo Jianmin

Lipid cytochemistry has not yet advanced far at the EM level. A major problem has been the loss of lipid during dehydration and embedding. Although the adoption of glutaraldehyde and osmium tetroxide accelerate the chemical reaction of lipid and osmium tetroxide can react on the double bouds of unsaturated lipid to from the osmium black, osmium tetroxide can be reduced in saturated lipid and subsequently some of unsaturated lipid are lost during dehydration. In order to reduce the loss of lipid by traditional method, some researchers adopted a few new methods, such as the change of embedding procedure and the adoption of new embedding media, to solve the problem. In a sense, these new methods are effective. They, however, usually require a long period of preparation. In this paper, we do research on the fiora nectary strucure of lauraceae by the rapid-embedding method wwith PEG under electron microscope and attempt to find a better method to solve the problem mentioned above.


2008 ◽  
Vol 42 (2) ◽  
pp. 48
Author(s):  
DOUG BRUNK

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