Wheat germ agglutinin chromatography of GlcNacβ1-3(GlcNAcβl-6)Gal and GlcNAcβ1-3(GlcNAcβ1-6)Galβ1-4GlcNAc, obtained by in vitro synthesis and by partial cleavage of teratocarcinoma poly-N-acetyllactosaminoglycans

1990 ◽  
Vol 68 (1) ◽  
pp. 44-53 ◽  
Author(s):  
Antti Seppo ◽  
Leena Penttilä ◽  
Anne Makkonen ◽  
Anne Leppänen ◽  
Ritva Niemelä ◽  
...  
Keyword(s):  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Embryonal Carcinoma ◽  
Periodate Oxidation ◽  
Carcinoma Cells ◽  
Teratocarcinoma Cell ◽  
Partial Acid Hydrolysis ◽  
In Vitro Synthesis ◽  
In Vitro Biosynthesis

GlcNAcβ1-3(GlcNAcβ1-6)[14C(U)]Gal and GlcNAcβ1-3(GlcNAcβ1-6)[14C(U)]Galβ1-4GlcNAc were prepared by in vitro synthesis. They were characterized by enzymatic sequencing, by partial acid hydrolysis, and by periodate oxidation experiments. The two saccharides were isolated also from partial acid hydrolysates of metabolically labeled poly-N- acetyllactosaminoglycans of murine embryonal carcinoma cells (line PC 13). The tetrasaccharide was retarded in a column of agarose-linked wheat germ agglutinin; the trisaccharide was strongly bound. Chromatography in this column separated the trisaccharide into two distinct peaks, which represented interconvertible molecules. Together with our previous data on linear teratocarcinoma saccharides, these findings show that affinity chromatography with immobilized wheat germ agglutinin can be advantageously used in fractionating radiolabeled oligo-N-acetyllactosaminoglycans and saccharides related to them.Key words: GlcNAcβ1-3(GlcNAcβ1-6)Gal, GlcNAcβ1-3(GlcNAcβ1-6)Galβ1-4GlcNAc, wheat germ agglutinin – agarose chromatography, in vitro biosynthesis, teratocarcinoma cell.

Immobilized wheat germ agglutinin separates small oligosaccharides derived from poly-N-acetyllactosaminoglycans of embryonal carcinoma cells

1988 ◽  
Vol 66 (5) ◽  
pp. 449-453 ◽  
Author(s):  
Ossi Renkonen ◽  
Pekka Mäkinen ◽  
Karl Hård ◽  
Jari Helin ◽  
Leena Penttilä
Keyword(s):  
Chromatographic Separation ◽  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Binding Affinities ◽  
Embryonal Carcinoma Cells ◽  
Small Column ◽  
Teratocarcinoma Cells ◽  
Weak Binding

Five pure oligosaccharides derived from poly-N-acetyllactosaminoglycans of teratocarcinoma cells were chromatographed on immobilized wheat germ agglutinin (WGA). Three of them, Galβ1-4GlcNAc, GlcNAcβ1-3Gal, and GlcNAcβ1-3Galβ1-4GlcNAc, revealed only weak binding, but GlcNAcβ1-6Galβ1-4GlcNAc was bound moderately and GlcNAcβ1-6Gal was bound quite strongly. The differences in the binding affinities were large enough to allow chromatographic separation of the five oligosaccharides into three distinct fractions in a small column of WGA–agarose.

scholarly journals Apoptosis of Human Embryonal Carcinoma Cells with in vitro Differentiation.

1996 ◽  
Vol 21 (1) ◽  
pp. 53-61 ◽  
Author(s):  
Taketo Yamada ◽  
Nao Suzuki ◽  
Nobuyoshi Hiraoka ◽  
Kentaroh Matsuoka ◽  
Sachiko Fukushima ◽  
...  
Keyword(s):  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
In Vitro Differentiation

scholarly journals Demethylation of somatic and testis-specific histone H2A and H2B genes in F9 embryonal carcinoma cells

1993 ◽  
Vol 13 (9) ◽  
pp. 5538-5548
Author(s):  
Y C Choi ◽  
C B Chae
Keyword(s):  
Embryonal Carcinoma ◽  
Cpg Island ◽  
Cpg Islands ◽  
High Density ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Cpg Dinucleotides ◽  
F9 Embryonal Carcinoma Cells ◽  
Methylation Patterns

In contrast to many other genes containing a CpG island, the testis-specific H2B (TH2B) histone gene exhibits tissue-specific methylation patterns in correlation with gene activity. Characterization of the methylation patterns within a 20-kb segment containing the TH2A and TH2B genes in comparison with that in a somatic histone cluster revealed that: (i) the germ cell-specific unmethylated domain of the TH2A and TH2B genes is defined as a small region surrounding the CpG islands of the TH2A and TH2B genes and (ii) somatic histone genes are unmethylated in both liver and germ cells, like other genes containing CpG islands, whereas flanking sequences are methylated. Transfection of in vitro-methylated TH2B, somatic H2B, and mouse metallothionein I constructs into F9 embryonal carcinoma cells revealed that the CpG islands of the TH2A and TH2B genes were demethylated like those of the somatic H2A and H2B genes and the metallothionein I gene. The demethylation of those CpG islands became significantly inefficient at a high number of integrated copies and a high density of methylated CpG dinucleotides. In contrast, three sites in the somatic histone cluster, of which two sites are located in the long terminal repeat of an endogenous retrovirus-like sequence, were efficiently demethylated even at a high copy number and a high density of methylated CpG dinucleotides. These results suggest two possible mechanisms for demethylation in F9 cells and methylation of CpG islands of the TH2A and TH2B genes at the postblastula stage during embryogenesis.

scholarly journals Trichomonas vaginalis Lipophosphoglycan Mutants Have Reduced Adherence and Cytotoxicity to Human Ectocervical Cells

2005 ◽  
Vol 4 (11) ◽  
pp. 1951-1958 ◽  
Author(s):  
Felix D. Bastida-Corcuera ◽  
Cheryl Y. Okumura ◽  
Angie Colocoussi ◽  
Patricia J. Johnson
Keyword(s):  
Wheat Germ Agglutinin ◽  
Parent Strain ◽  
Trichomonas Vaginalis ◽  
Wheat Germ ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Host Cells ◽  
Chemical Mutagenesis ◽  
Reduced Adherence

ABSTRACT The extracellular human pathogen Trichomonas vaginalis is covered by a dense glycocalyx thought to play a role in host-parasite interactions. The main component of the glycocalyx is lipophosphoglycan (LPG), a polysaccharide anchored in the plasma membrane by inositol phosphoceramide. To study the role of LPG in trichomonads, we produced T. vaginalis LPG mutants by chemical mutagenesis and lectin selection and characterized them using morphological, biochemical, and functional assays. Two independently selected LPG mutants, with growth rates comparable to that of the wild-type (parent) strain, lost the ability to bind the lectins Ricinnus comunis agglutinin I (RCA120) and wheat germ agglutinin, indicating alterations in surface galactose and glucosamine residues. LPG isolated from mutants migrated faster than parent strain LPG on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, suggesting the mutants had shorter LPG molecules. Dionex high-performance anion exchange chromatography with pulsed amperometric detection analyses revealed galactosamine, glucosamine, galactose, glucose, mannose/xylose, and rhamnose as the main monosaccharides of T. vaginalis parent strain LPG. LPG from both mutants showed a reduction of galactose and glucosamine, corresponding with the reduced size of their LPG molecules and inability to bind the lectins RCA120 and wheat germ agglutinin. Mutant parasites were defective in attachment to plastic, a characteristic associated with avirulent strains of T. vaginalis. Moreover, the mutants were less adherent and less cytotoxic to human vaginal ectocervical cells in vitro than the parental strain. Finally, while parent strain LPG could inhibit the attachment of parent strain parasites to vaginal cells, LPG from either mutant could not inhibit attachment. These combined results demonstrate that T. vaginalis adherence to host cells is LPG mediated and that an altered LPG leads to reduced adherence and cytotoxicity of this parasite.

Differential effects of epidermal growth factor (EGF) on cell locomotion and cell proliferation in a cloned human embryonal carcinoma-derived cell line in vitro

1986 ◽  
Vol 86 (1) ◽  
pp. 47-55
Author(s):  
W. Engstrom
Keyword(s):  
Cell Proliferation ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Cell Line ◽  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Colony Diameter ◽  
Cell Locomotion ◽  
Epidermal Growth

The effects of epidermal growth factor (EGF) on clones from a human embryonal carcinoma-derived cell line (Tera-2) have been studied. Cells were plated at clonal densities, whereafter the effects of serum and EGF on cell locomotion and cell proliferation were examined. The addition of 50 ngEGF ml-1 resulted in increased migration, as judged by increased colony diameter in the presence of EGF. However, the effect of EGF on cell locomotion was rarely accompanied by any effect on cell proliferation. It was concluded that EGF exerts a preferential effect on cell migration in human embryonal carcinoma cells in vitro.

scholarly journals Demethylation of somatic and testis-specific histone H2A and H2B genes in F9 embryonal carcinoma cells.

1993 ◽  
Vol 13 (9) ◽  
pp. 5538-5548 ◽  
Author(s):  
Y C Choi ◽  
C B Chae
Keyword(s):  
Embryonal Carcinoma ◽  
Cpg Island ◽  
Cpg Islands ◽  
High Density ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Cpg Dinucleotides ◽  
F9 Embryonal Carcinoma Cells ◽  
Methylation Patterns

In contrast to many other genes containing a CpG island, the testis-specific H2B (TH2B) histone gene exhibits tissue-specific methylation patterns in correlation with gene activity. Characterization of the methylation patterns within a 20-kb segment containing the TH2A and TH2B genes in comparison with that in a somatic histone cluster revealed that: (i) the germ cell-specific unmethylated domain of the TH2A and TH2B genes is defined as a small region surrounding the CpG islands of the TH2A and TH2B genes and (ii) somatic histone genes are unmethylated in both liver and germ cells, like other genes containing CpG islands, whereas flanking sequences are methylated. Transfection of in vitro-methylated TH2B, somatic H2B, and mouse metallothionein I constructs into F9 embryonal carcinoma cells revealed that the CpG islands of the TH2A and TH2B genes were demethylated like those of the somatic H2A and H2B genes and the metallothionein I gene. The demethylation of those CpG islands became significantly inefficient at a high number of integrated copies and a high density of methylated CpG dinucleotides. In contrast, three sites in the somatic histone cluster, of which two sites are located in the long terminal repeat of an endogenous retrovirus-like sequence, were efficiently demethylated even at a high copy number and a high density of methylated CpG dinucleotides. These results suggest two possible mechanisms for demethylation in F9 cells and methylation of CpG islands of the TH2A and TH2B genes at the postblastula stage during embryogenesis.

scholarly journals Ectopic activation of WNT signaling in human embryonal carcinoma cells and its effects in short- and long-term in vitro culture

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yaser Atlasi ◽  
Rebecca T. van Dorsten ◽  
Andrea Sacchetti ◽  
Rosalie Joosten ◽  
J. Wolter Oosterhuis ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
In Vitro Culture ◽  
Embryonal Carcinoma ◽  
Carcinoma Cells ◽  
Embryonal Carcinoma Cells ◽  
Ectopic Activation ◽  
Short And Long Term

Wheat germ agglutinin-grafted lipid nanoparticles: Preparation and in vitro evaluation of the association with Caco-2 monolayers

2010 ◽  
Vol 397 (1-2) ◽  
pp. 155-163 ◽  
Author(s):  
Ying Liu ◽  
Pengfei Wang ◽  
Chen Sun ◽  
Nianping Feng ◽  
Wuxiong Zhou ◽  
...  
Keyword(s):  
Wheat Germ Agglutinin ◽  
Wheat Germ ◽  
Lipid Nanoparticles ◽  
In Vitro Evaluation
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