Analysis of protein patterns in two-dimensional gels of cultured human cells with trisomy 21.

1982 ◽  
Vol 28 (4) ◽  
pp. 987-992 ◽  
Author(s):  
J Klose ◽  
E Zeindl ◽  
K Sperling
Keyword(s):  
Cell Lines ◽  
Fetal Lung ◽  
Staining Intensity ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Cell Clones ◽  
Cultured Human Cells ◽  
Regulator Genes ◽  
Control Patterns ◽  
Trisomic Cell

Abstract The occurrence of one chromosome of the cell in triplicate (trisomy, Ts) should increase the amount of all cell proteins coded by genes located on this chromosome. Many other proteins should be altered in their quantity by regulator genes, present in a threefold dosage, and by several indirect effects of the trisomy. We used two-dimensional electrophoresis to investigate the effect of the human Ts 21 on the proteins. Cells from different individuals with Ts 21 were cultured. Seven cell lines were derived from skin tissue and four from cells in amniotic fluid, and two cell clones were raised from fetal lung with Ts 21 mosaic. Stained two-dimensional protein patterns from these cell lines were compared with control patterns, and clearly visible differences in the staining intensity of corresponding polypeptide spots were evaluated. A few quantitatively variant polypeptides occurred in all trisomic cell lines investigated that were of a particular cell type, but no variants were consistently present in all of the 13 cell lines investigated. However the total number of variants (including variants not found in all individual cell lines) was considerably higher in trisomic cells than in normal cells.

Chromosomes and sex differentiation in eutherians

Development ◽  
1984 ◽  
Vol 83 (Supplement) ◽  
pp. 41-49
Author(s):  
Ulrich Müller
Keyword(s):  
Developmental Stages ◽  
Sex Chromosome ◽  
Sex Differentiation ◽  
Gonadal Development ◽  
Relative Molecular Mass ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Two Dimensional Gel Electrophoresis ◽  
Cell Clones ◽  
Specific Proteins

In order to learn more about the role of sex chromosome-dependent gene products in gonadogenesis, changes in protein patterns were studied during gonadal development. Two-dimensional gel electrophoresis analysis revealed specific proteins in both sexes at all developmental stages. Evidently the gonads are not indifferent by biochemical criteria at any developmental stage and express several specific genes from the onset of differentiation. To correlate these polypeptides with the sex chromosomes, proteins were investigated in human–rodent somatic cell hybrids and in genetically identical cell clones differing in one sex chromosome only. On two-dimensional gels one Y-dependent polypeptide was found with similar characteristics (relative molecular mass and isoelectric point) as an early testicular polypeptide. Its identity, however, remains to be proven.

(EB) Virus: Latency and Expression in Transplanted and Cultured Human Cells

1971 ◽  
Vol 29 ◽  
pp. 368-369
Author(s):  
B. G. Uzman ◽  
M. M. Kasac ◽  
H. Saito ◽  
A. Krishan
Keyword(s):  
Cell Lines ◽  
Primary Cultures ◽  
Virus Particles ◽  
Barr Virus ◽  
Virus Latency ◽  
Virus Surveillance ◽  
Cultured Human Cells ◽  
Epstein Barr ◽  
Serial Transplantation ◽  
Tubular Arrays

In conjunction with the cultivation and transplantation of cells from human tumors by the Programs of Microbiology and Immunogenetics, virus surveillance by electron microscopy has been routinely employed. Of particular interest in this regard have been 3 cell lines cultured from lymph nodes or spleen of 2 patients with Hodgkin's disease and 1 patient with Letterer-Siwe's disease. Each of these cell lines when transplanted in Syrian hamster neonates conditioned with anti-lymphocyte serum grew as serially transplantable tumors; from such transplants of the 3 cell lines cell cultures were retrieved.Herpes type virus particles (Figs. 1, 2, 3) were found in the primary cultures of all three lines, in frozen thawed aliquots of same, and in cultures retrieved from their tumors growing by serial transplantation in hamsters. No virus was detected in sections of 25 of the serially transplanted tumors. However, in 10 such tumors there were repeated instances of tubular arrays in the cisternae of the endoplasmic reticulum (Fig. 4). On serologic examination the herpes virus was shown to be the Epstein-Barr virus.

scholarly journals BHK-21 Cell Clones Differ in Chikungunya Virus Infection and MXRA8 Receptor Expression

Viruses ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 949
Author(s):  
Peiqi Yin ◽  
Margaret Kielian
Keyword(s):  
Cell Lines ◽  
Primary Infection ◽  
Sindbis Virus ◽  
Chikungunya Virus ◽  
Mrna Level ◽  
Receptor Expression ◽  
Chikv Infection ◽  
Cell Clones ◽  
Clonal Cell Lines ◽  
Exogenous Expression

Baby hamster kidney-21 (BHK-21) cells are widely used to propagate and study many animal viruses using infection and transfection techniques. Among various BHK-21 cell clones, the fibroblast-like BHK-21/C-13 line and the epithelial-like BHK-21/WI-2 line are commonly used cell clones for alphavirus research. Here we report that BHK-21/WI-2 cells were significantly less susceptible to primary infection by the alphavirus chikungunya virus (CHIKV) than were BHK-21/C-13 cells. The electroporation efficiency of alphavirus RNA into BHK-21/WI-2 was also lower than that of BHK-21/C-13. The growth of CHIKV was decreased in BHK-21/WI-2 compared to BHK-21/C-13, while primary infection and growth of the alphavirus Sindbis virus (SINV) were equivalent in the two cell lines. Our results suggested that CHIKV entry could be compromised in BHK-21/WI-2. Indeed, we found that the mRNA level of the CHIKV receptor MXRA8 in BHK-21/WI-2 cells was much lower than that in BHK-21/C-13 cells, and exogenous expression of either human MXRA8 or hamster MXRA8 rescued CHIKV infection. Our results affirm the importance of the MXRA8 receptor for CHIKV infection, and document differences in its expression in two clonal cell lines derived from the original BHK-21 cell cultures. Our results also indicate that CHIKV propagation and entry studies in BHK-21 cells will be significantly more efficient in BHK-21/C-13 than in BHK-21/WI-2 cells.

scholarly journals The plasminogen system and cell surfaces: evidence for plasminogen and urokinase receptors on the same cell type.

1986 ◽  
Vol 103 (6) ◽  
pp. 2411-2420 ◽  
Author(s):  
E F Plow ◽  
D E Freaney ◽  
J Plescia ◽  
L A Miles
Keyword(s):  
Cell Line ◽  
Plasminogen Activator ◽  
Cell Lines ◽  
Specific Binding ◽  
High Capacity ◽  
Fetal Lung ◽  
Fibroblast Cell ◽  
Cell Types ◽  
Cell Surfaces ◽  
Catalytically Active

The capacity of cells to interact with the plasminogen activator, urokinase, and the zymogen, plasminogen, was assessed using the promyeloid leukemic U937 cell line and the diploid fetal lung GM1380 fibroblast cell line. Urokinase bound to both cell lines in a time-dependent, specific, and saturable manner (Kd = 0.8-2.0 nM). An active catalytic site was not required for urokinase binding to the cells, and 55,000-mol-wt urokinase was selectively recognized. Plasminogen also bound to the two cell lines in a specific and saturable manner. This interaction occurred with a Kd of 0.8-0.9 microM and was of very high capacity (1.6-3.1 X 10(7) molecules bound/cell). The interaction of plasminogen with both cell types was partially sensitive to trypsinization of the cells and required an unoccupied high affinity lysine-binding site in the ligand. When plasminogen was added to the GM1380 cells, a line with high intrinsic plasminogen activator activity, the bound ligand was comprised of both plasminogen and plasmin. Urokinase, in catalytically active or inactive form, enhanced plasminogen binding to the two cell lines by 1.4-3.3-fold. Plasmin was the predominant form of the bound ligand when active urokinase was added, and preformed plasmin can also bind directly to the cells. Plasmin on the cell surface was also protected from its primary inhibitor, alpha 2-antiplasmin. These results indicate that the two cell lines possess specific binding sites for plasminogen and urokinase, and a family of widely distributed cellular receptors for these components may be considered. Endogenous or exogenous plasminogen activators can generate plasmin on cell surfaces, and such activation may provide a mechanism for arming cell surfaces with the broad proteolytic activity of this enzyme.

scholarly journals Protein expression profile related to cisplatin resistance in bladder cancer cell lines detected by two-dimensional gel electrophoresis

2015 ◽  
Vol 36 (4) ◽  
pp. 253-261 ◽  
Author(s):  
Yoshinori TAOKA ◽  
Kazumasa MATSUMOTO ◽  
Kazuya OHASHI ◽  
Satoru MINAMIDA ◽  
Masahiro HAGIWARA ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Protein Expression ◽  
Cell Lines ◽  
Cancer Cell ◽  
Expression Profile ◽  
Cisplatin Resistance ◽  
Cancer Cell Lines ◽  
Bladder Cancer Cell ◽  
Two Dimensional ◽  
Protein Expression Profile

Fabrication and Cytotoxicity Studies of the TiO2 Doped with Copper-Based Nanocomposite Particles

2011 ◽  
Vol 695 ◽  
pp. 393-396 ◽  
Author(s):  
Chang Mao Hung
Keyword(s):  
Fetal Lung ◽  
Copper Nitrate ◽  
Tissue Cell ◽  
Nanocomposite Particles ◽  
Toxicological Effect ◽  
Cytotoxicity Studies ◽  
Cultured Human Cells ◽  
Cytotoxicity Assessment ◽  
Human Fetal Lung ◽  
Dispersion Character

Since the growing interest in the manufacture and environmental applications of nanocomposites consisting of CuO and TiO2nanoparticles (NPs), related toxicological effect and interaction with cellular structures for these newly developed materials are still unknown. Recent literature reveals that nanosized CuO and TiO2particles have cytotoxicity risks and ability to cause oxidative stress on health. This work considers the CuO doped TiO2nanocomposite particles were synthesized via a coprecipitation method with aqueous solutions as precursors of copper nitrateand titanium dioxide. Moreover, the nanocomposite particles were characterized using TGA-DTA, UV-Vis and TEM measurements. The calcination temperature was selected at 873 K. The nanocomposite particles were characterized by TEM, as the primary particles, aggregates ranged from 30 to 100 nm and have a good dispersion character. Cell cytotoxicity assessment and the percentage cell survival was determined by using 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethoxyphenol)-2-(4-sulfophenyl)-2H-tetrazoli um (MTS) assay on human fetal lung tissue cell (MRC-5). The experimental results show that the CuO doped TiO2nanocomposite particles cause potential cytotoxicity effect in cultured human cells.

scholarly journals Tissue type-specific expression of intermediate filament proteins in a cultured epithelial cell line from bovine mammary gland

1983 ◽  
Vol 96 (1) ◽  
pp. 37-50 ◽  
Author(s):  
E Schmid ◽  
DL Schiller ◽  
C Grund ◽  
J Stadler ◽  
WW Franke
Keyword(s):  
Mammary Gland ◽  
Epithelial Cell ◽  
Cell Line ◽  
Cell Lines ◽  
Intermediate Filament ◽  
Striking Difference ◽  
Intermediate Filament Proteins ◽  
Cell Clones ◽  
Vimentin Filaments

Different clonal cell lines have been isolated from cultures of mammary gland epithelium of lactating cow's udder and have been grown in culture media containing high concentrations of hydrocortisone, insulin, and prolactin. These cell (BMGE+H), which grow in monolayers of typical epithelial appearance, are not tightly packed, but leave intercellular spaces spanned by desmosomal bridges. The cells contain extended arrays of cytokeratin fibrils, arranged in bundles attached to desmosomes. Gel electophoresis show that they synthesize cytokeratins similar, if not identical, to those found in bovine epidermis and udder, including two large (mol wt 58,500 and 59,000) and basic (pH range: 7-8) and two small (mol wt 45,500 and 50,000) and acidic (pH 5.32 and 5.36) components that also occur in phosphorylated forms. Two further cytokeratins of mol wts 44,000 (approximately pH 5.7) and 53,000 (pH 6.3) are detected as minor cytokeratins in some cell clones. BMGE+H cells do not produce vimentin filaments as determined by immunofluorescence microscopy and gel electrophoresis. By contrast, BMGE-H cells, which have emerged from the same original culture but have been grown without hormones added, are not only morphologically different, but also contain vimentin filaments and a different set of cytokeratins, the most striking difference being the absence of the two acidic cytokeratins of mol wt 50,000 and 45,500. Cells of the BMGE+H line are characterized by an unusual epithelial morphology and represent the first example of a nonmalignant permanent cell line in vitro that produces cytokeratin but not vimentin filaments. The results show that (a) tissue-specific patterns of intermediate filament expression can be maintained in permanent epithelial cell lines in culture, at least under certain growth conditions; (b) loss of expression of relatively large, basic cytokeratins is not an inevitable consequence of growth of epithelial cells in vitro. Our results further show that, during culturing, different cell clones with different cytoskeletal composition can emerge from the same cell population and suggest that the presence of certain hormones may have an influence on the expression of intermediate filament proteins.

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