scholarly journals Murine Gammaherpesvirus (MHV-68) Transforms Cultured Cells in vitro

Intervirology ◽  
2015 ◽  
Vol 58 (2) ◽  
pp. 69-72 ◽  
Author(s):  
Veronika Mrázová ◽  
Tatiana Betáková ◽  
Marcela Kúdelová ◽  
Miroslava Šupolíková ◽  
Veronika Lachová ◽  
...  
Keyword(s):  
Cell Lines ◽  
Cultured Cells ◽  
Virus Antigen ◽  
Immunofluorescence Assay ◽  
Dermal Fibroblasts ◽  
Murine Gammaherpesvirus ◽  
Transformed Cell ◽  
Transformed Cell Lines ◽  
Nih 3T3

Human dermal fibroblasts and mouse NIH/3T3 cells acquired the transformed phenotype (‘criss-cross' pattern of growth) after infection with ultraviolet-irradiated murine gammaherpesvirus (MuHV-4 strain 68; MHV-68). These cells with changed phenotype could be serially cultured for 5-6 passages (35-40 days), and then they entered into crisis and most of them died. In a small number of cultures, however, foci of newly transformed cells appeared from which two stable cell lines were derived. After 6-9 cell culture passages of the MHV-68 transformed cell lines, MHV-68 DNA and virus antigen could be detected by PCR and immunofluorescence assay along with the disappearance of actin bundles, indicating that both transformed cell lines might be oncogenic.

scholarly journals Effects of Physicochemical Properties of Polyacrylamide (PAA) and Polydimethylsiloxane (PDMS) on Cardiac Cell behavior

Soft Matter ◽  
2021 ◽  
Author(s):  
Karim Daliri ◽  
Kurt Pfannkuche ◽  
Bora Garipcan
Keyword(s):  
Cell Lines ◽  
Cultured Cells ◽  
Cell Behavior ◽  
Cardiac Cell ◽  
Established Cell Lines ◽  
The World ◽  
Established Cell ◽  
Primary Origin ◽  
Transformed Cell Lines

In vitro cell culture is commonly applied in laboratories around the world. Cultured cells are either of primary origin or established cell lines. Such transformed cell lines are increasingly replaced...

The antiproliferative effects of agmatine correlate with the rate of cellular proliferation

2007 ◽  
Vol 293 (2) ◽  
pp. C705-C711 ◽  
Author(s):  
Masato Isome ◽  
Mark J. Lortie ◽  
Yasuko Murakami ◽  
Eva Parisi ◽  
Senya Matsufuji ◽  
...  
Keyword(s):  
Cell Line ◽  
Cell Lines ◽  
Cellular Proliferation ◽  
Intracellular Accumulation ◽  
3T3 Cells ◽  
Antiproliferative Effects ◽  
Transformed Cell ◽  
Polyamine Transport ◽  
Transformed Cell Lines ◽  
Nih 3T3

Polyamines are small cationic molecules required for cellular proliferation. Agmatine is a biogenic amine unique in its capacity to arrest proliferation in cell lines by depleting intracellular polyamine levels. We previously demonstrated that agmatine enters mammalian cells via the polyamine transport system. As polyamine transport is positively correlated with the rate of cellular proliferation, the current study examines the antiproliferative effects of agmatine on cells with varying proliferative kinetics. Herein, we evaluate agmatine transport, intracellular accumulation, and its effects on antizyme expression and cellular proliferation in nontransformed cell lines and their transformed variants. H-ras- and Src-transformed murine NIH/3T3 cells (Ras/3T3 and Src/3T3, respectively) that were exposed to exogenous agmatine exhibit increased uptake and intracellular accumulation relative to the parental NIH/3T3 cell line. Similar increases were obtained for human primary foreskin fibroblasts relative to a human fibrosarcoma cell line, HT1080. Agmatine increases expression of antizyme, a protein that inhibits polyamine biosynthesis and transport. Ras/3T3 and Src/3T3 cells demonstrated augmented increases in antizyme protein expression relative to NIH/3T3 in response to agmatine. All transformed cell lines were significantly more sensitive to the antiproliferative effects of agmatine than nontransformed lines. These effects were attenuated in the presence of exogenous polyamines or inhibitors of polyamine transport. In conclusion, the antiproliferative effects of agmatine preferentially target transformed cell lines due to the increased agmatine uptake exhibited by cells with short cycling times.

scholarly journals Generation of tumorigenic porcine pancreatic ductal epithelial cells: toward a large animal model of pancreatic cancer

2018 ◽  
Author(s):  
Neeley Remmers ◽  
Jesse L. Cox ◽  
James A. Grunkemeyer ◽  
Shruthi Aravind ◽  
Christopher K. Arkfeld ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Animal Model ◽  
Cell Lines ◽  
Large Animal Model ◽  
Large Animal ◽  
Murine Models ◽  
Transformed Cell ◽  
Transformed Cell Lines

AbstractBackground. A large animal model of pancreatic cancer would permit development of diagnostic and interventional technologies not possible in murine models, and also would provide a more biologically-relevant platform for penultimate testing of novel therapies, prior to human testing. Here, we describe our initial studies in the development of an autochthonous, genetically-defined, large animal model of pancreatic cancer, using immunocompetent pigs.Methods. Primary pancreatic epithelial cells were isolated from pancreatic duct of domestic pigs; epithelial origin was confirmed with immunohistochemistry. Three transformed cell lines subsequently were generated from these primary cells using expression of oncogenic KRAS and dominant negative p53, with/without knockdown of p16 and SMAD4. We tested these cell lines using in vitro and in vivo assays of transformation and tumorigenesis.Results. The transformed cell lines outperformed the primary cells in terms proliferation, population doubling time, soft agar growth, 2D migration, and Matrigel invasion, with the greatest differences observed when all four genes (KRAS, p53, p16, and SMAD4) were targeted. All three transformed cell lines grew tumors when injected subcutaneously in nude mice, demonstrating undifferentiated morphology, mild desmoplasia, and staining for both epithelial and mesenchymal markers. Injection into the pancreas of nude mice resulted in distant metastases, particularly when all four genes were targeted.Conclusions. Tumorigenic porcine pancreatic cell lines were generated. Inclusion of four genetic “hits” (KRAS, p53, p16, and SMAD4) appeared to produce the best results in our in vitro and in vivo assays. The next step will be to perform autologous or syngeneic implantation of these cell lines into the pancreas of immunocompetent pigs. We believe that the resultant large animal model of pancreatic cancer could supplement existing murine models, thus improving preclinical research on diagnostic, interventional, and therapeutic technologies.

In vitro traits of adenovirus-transformed cell lines and their relevance to tumorigenicity in nude mice

Cell ◽  
1977 ◽  
Vol 10 (4) ◽  
pp. 669-678 ◽  
Author(s):  
Phillip H. Gallimore ◽  
James K. McDougall ◽  
Lan Bo Chen
Keyword(s):  
Cell Lines ◽  
Nude Mice ◽  
Transformed Cell ◽  
Transformed Cell Lines

scholarly journals Activation and association of Stat3 with Src in v-Src-transformed cell lines.

1996 ◽  
Vol 16 (4) ◽  
pp. 1595-1603 ◽  
Author(s):  
X Cao ◽  
A Tay ◽  
G R Guy ◽  
Y H Tan
Keyword(s):  
Growth Factor ◽  
Cell Lines ◽  
Tyrosine Kinases ◽  
New Members ◽  
Signal Transducers ◽  
Stat Proteins ◽  
Transformed Cell ◽  
Transformed Cell Lines

STAT proteins are a group of latent cytoplasmic transcription factors which function as signal transducers and activators of transcription. Stat1 and -2 were originally identified to function in interferon signaling, and Stat1 was also found to be activated by epidermal growth factor (EGF) and other cytokines. New members of the STAT gene family are identified. Among them, Stat3 has 52.5% amino acid sequence homology with Stat1 and is activated by platelet-derived growth factor (PDGF), colony-stimulating factor 1 (CSF-1), EGF, interleukin-6, and other cytokines. Treatment of cells with EGF activates Stat1 and Stat3, which become phosphorylated on tyrosine residues to form homo - or heterodimers and translocate into the nucleus, binding to the sis-inducible element (SIE) in the c-fos promoter. Somatic cell genetic analyses demonstrated that Jaks, a family of nontransmembrane protein tyrosine kinases, are required for the activation of Stat1 and Stat2 in interferon-treated cells. However, little is known about the activation of Stat3 by growth factors. Here we report that in all v-Src-transformed cell lines examined, Stat3 is constitutively activated to bind to DNA and the phosphorylation of tyrosine on Stat3 is enhanced by the induction of v-Src expression. We also report that Src is shown to be associated with Stat3 in vivo, as well as in vitro, and phosphorylates Stat3 in vitro. Stat3 is also activated by CSF-1, possibly through CSF-1 receptor-c Src association in NIH 3T3 cells overexpressing CSF-1 receptors. Together, the data suggest that Src is involved in activation of Stat3 in growth factor signal transduction.

Rapid preparation of nucleotides from acid-soluble pools by chromatography on silica, as exemplified with acid extracts of cultured cells.

1980 ◽  
Vol 26 (10) ◽  
pp. 1430-1434 ◽  
Author(s):  
C D Lothrop ◽  
M Uziel
Keyword(s):  
Nucleic Acid ◽  
Cell Lines ◽  
Cultured Cells ◽  
Silica Gels ◽  
Rapid Preparation ◽  
Rapid Technique ◽  
Transformed Cell ◽  
Water Solvent ◽  
Retention Volumes ◽  
Transformed Cell Lines

Abstract A rapid technique (5-10 min) has been developed for fractionating nucleotides from base and nucleoside contaminants in acid extracts of cells, by adsorption to silica gels. Silica gels (1-mL bed volume) were washed with 5 mL of water then with 5 mL of acetonitrile/water (90/10 by vol). After applying 3-mL samples, adjusted to 900 mL/L acetonitrile content, we washed the gel with an additional 10 mL of the acetonitrile/water solvent. More than 95% of the amounts of bases and nucleosides prsent, except for cytidine (92%), did not adsorb to silica under these conditions. Nucleotides were then quantitatively eluted with 9 mL of water. The retention volumes for positive, negative, and neutral nucleic acid components have been determined, to investigate the discriminatory properties of nucleic acid components on silica. Compounds (bases, nucleosides) that are not ionized at pH 7 do not bind to silica. However, negative, positive, and zwitterionic compounds are tightly adsorbed to the silica gels. This procedure has been used to purify nucleotides from several normal and transformed cell lines.

scholarly journals Spi-1/PU.1 Is a Positive Regulator of the Fli-1 Gene Involved in Inhibition of Erythroid Differentiation in Friend Erythroleukemic Cell Lines

1999 ◽  
Vol 19 (1) ◽  
pp. 121-135 ◽  
Author(s):  
Joëlle Starck ◽  
Alexandre Doubeikovski ◽  
Sandrine Sarrazin ◽  
Colette Gonnet ◽  
Govinda Rao ◽  
...  
Keyword(s):  
Cell Lines ◽  
Leukemia Virus ◽  
Erythroid Differentiation ◽  
Specific Pattern ◽  
Expression Vectors ◽  
Common Mechanism ◽  
Transformed Cell ◽  
Erythroleukemic Cell ◽  
Transformed Cell Lines

ABSTRACT Spi-1/PU.1 and Fli-1 are two members of the ETS family of transcription factors whose expression is deregulated by proviral insertion in most erythroleukemic cell lines induced by the spleen focus-forming virus (SFFV) and Friend murine leukemia virus (F-MuLV) components of the Friend viral complex, respectively. In this study, we present evidence that transcription of the Fli-1 gene is positively regulated by Spi-1/PU.1 in SFFV-transformed cell lines: (i) all SFFV-transformed cell lines expressing Spi-1/PU.1 are characterized by a specific pattern of Fli-1 gene transcripts initiated in the −200 region instead of position −400 as reported for F-MuLV-transformed cell lines; (ii) these Fli-1 transcripts initiated in the −200 region are downregulated in parallel with that of Spi-1/PU.1 during hexamethylenebisacetamide (HMBA) induced differentiation; and (iii) Fli-1 transcription is upregulated in SFFV cells lines following stable transfection of a Spi-1/PU.1 expression vector. Furthermore, we found by transient transfection assays that the −270/−41 region of the Fli-1 gene displays promoter activity which is transactivated by Spi-1/PU.1. This promoter is strictly dependent on the integrity of two highly conserved ETS DNA binding sites that bind the Spi-1/PU.1 protein in vitro. Finally, we show that transfection of constitutive or inducible Fli-1 expression vectors in SFFV-transformed cells inhibits their erythroid differentiation induced by HMBA. Overall, these data indicate that Fli-1 is a target gene of the Spi-1/PU.1 transcription factor in SFFV-transformed cell lines. We further suggest that deregulated synthesis of Fli-1 may trigger a common mechanism contributing to erythroleukemia induced by either SFFV or F-MuLV.

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