Development of a mouse antiperoxidase secreting hybridoma for use in the production of a mouse PAP complex for immunocytochemistry and as a parent cell line in the development of hybrid hybridomas

1985 ◽  
Vol 83 (5) ◽  
pp. 405-408 ◽  
Author(s):  
F. M. Semenenko ◽  
S. Bramwell ◽  
E. Sidebottom ◽  
A. C. Cuello
Keyword(s):  
Cell Line ◽  
Parent Cell ◽  
Parent Cell Line

scholarly journals Involvement of APOBEC3B in mutation induction by irradiation

2020 ◽  
Vol 61 (6) ◽  
pp. 819-827
Author(s):  
Yohei Saito ◽  
Hiromasa Miura ◽  
Nozomi Takahashi ◽  
Yoshikazu Kuwahara ◽  
Yumi Yamamoto ◽  
...  
Keyword(s):  
Dna Damage ◽  
Cancer Risk ◽  
Cell Line ◽  
Human Cancer ◽  
Spontaneous Mutation ◽  
Mutation Induction ◽  
Parent Cell ◽  
Induced Mutations ◽  
Parent Cell Line ◽  
Radiation Induced

Abstract To better understand the cancer risk posed by radiation and the development of radiation therapy resistant cancer cells, we investigated the involvement of the cancer risk factor, APOBEC3B, in the generation of radiation-induced mutations. Expression of APOBEC3B in response to irradiation was determined in three human cancer cell lines by real-time quantitative PCR. Using the hypoxanthine-guanine phosphoribosyl transferase (HPRT) mutation assay, mutations in the HPRT gene caused by irradiation were compared between APOBEC3B-deficient human hepatocellular carcinoma (HepG2) cells [APOBEC3B knocked out (KO) using CRISPR-Cas9 genome editing] and the parent cell line. Then, HPRT-mutated cells were individually cultured to perform PCR and DNA sequencing of HPRT exons. X-Irradiation induced APOBEC3B expression in HepG2, human cervical cancer epithelial carcinoma (HeLa) and human oral squamous cell carcinoma (SAS) cells. Forced expression of APOBEC3B increased spontaneous mutations. By contrast, APOBEC3B KO not only decreased the spontaneous mutation rate, but also strongly suppressed the increase in mutation frequency after irradiation in the parent cell line. Although forced expression of APOBEC3B in the nucleus caused DNA damage, higher levels of APOBEC3B tended to reduce APOBEC3B-induced γ-H2AX foci formation (a measure of DNA damage repair). Further, the number of γ-H2AX foci in cells stably expressing APOBEC3B was not much higher than that in controls before and after irradiation, suggesting that a DNA repair pathway may be activated. This study demonstrates that irradiation induces sustained expression of APOBEC3B in HepG2, HeLa and SAS cells, and that APOBEC3B enhances radiation-induced partial deletions.

Generation of megakaryocytes and platelets from preadipocyte cell line 3T3-L1, but not the parent cell line 3T3, in vitro

2010 ◽  
Vol 402 (4) ◽  
pp. 796-800 ◽  
Author(s):  
Yumiko Matsubara ◽  
Hidenori Suzuki ◽  
Yasuo Ikeda ◽  
Mitsuru Murata
Keyword(s):  
Cell Line ◽  
Parent Cell ◽  
Parent Cell Line

scholarly journals Absorptive and mucus-secreting subclones isolated from a multipotent intestinal cell line (HT-29) provide new models for cell polarity and terminal differentiation.

1987 ◽  
Vol 105 (1) ◽  
pp. 345-357 ◽  
Author(s):  
C Huet ◽  
C Sahuquillo-Merino ◽  
E Coudrier ◽  
D Louvard
Keyword(s):  
Cell Line ◽  
Cell Population ◽  
Morphological Characteristics ◽  
Human Tumor ◽  
Intestinal Cell ◽  
Parent Cell ◽  
Absorptive Cells ◽  
Parent Cell Line ◽  
Ht 29

A clone HT29-18 has been isolated from the parent cell line HT-29, which derived from a human colon adenocarcinoma (Fogh, J., and G. Trempe, 1975, Human Tumor Cells in Vitro, J. Fogh, editor, Plenum Publishing Corp., New York, 115-141). This clone is able to differentiate as the parent cell line does. Differentiation occurs when glucose is replaced by galactose in the culture medium (Pinto, M., M.D. Appay, P. Simon-Assman, G. Chevalier, N. Dracopoli, J. Fogh, and A. Zweibaum, 1982, Biol. Cell., 44:193-196). We demonstrate here that the differentiated cloned population HT29-18/gal is heterogenous: although 90% of the cells show morphological characteristics of "absorptive cells", only 20-30% of them display sucrase-isomaltase in their apical microvillar membranes. About 10% of the entire cell population consists of cells containing mucous granules similar to intestinal goblet cells. We have isolated two subclones, HT29-18-C1 and HT29-18-N2, from the differentiated HT29-18/gal cells. HT29-18-C1 cells show morphological characteristics of polarized absorptive cells, when growing either in glucose- or in galactose-containing media, but the sucrase-isomaltase is not expressed in the cells grown in glucose-containing medium. The clone HT29-18-N2 is also polarized in both culture conditions and is similar to globlet cells in vivo. It grows as a monolayer, exhibits tight junctions, and contains numerous mucous granules whose exocytosis can be triggered by carbachol, a parasympathomimetic drug. We conclude that the clone HT29-18 first isolated was a multipotent cell population from which we isolated several subclones that differentiate either as absorptive (HT29-18-C1) or as mucous (HT29-18-N2) cells. In contrast to the parent HT-29 cell line, the subclones retain most of their differentiated properties in glucose-containing medium.

DNA rearrangement causes a high rate of spontaneous mutation at the immunoglobulin heavy-chain locus of a mouse myeloma cell line

1986 ◽  
Vol 6 (12) ◽  
pp. 4228-4235
Author(s):  
H Yu ◽  
L A Eckhardt
Keyword(s):  
Cell Line ◽  
Heavy Chain ◽  
Spontaneous Mutation ◽  
Immunoglobulin Heavy Chain ◽  
Myeloma Cell ◽  
Chain Gene ◽  
Parent Cell ◽  
Mrna Levels ◽  
Dna Rearrangement ◽  
Myeloma Cell Line

The spontaneous mutation rate of immunoglobulin genes expressed in myeloma cells is well above that of other genes expressed in these or in other cell types. The nature of such mutations in one myeloma cell line, MPC11, was explored at the molecular level. Included in this study were MPC11 variants representing 24 independent and spontaneous mutations affecting immunoglobulin secretion. Of the mutants studied, 19 had ceased immunoglobulin heavy chain (IgH) production (nonproducers), and 5 produced from as little as 1/1,000 to as much as 1/10 the amount of immunoglobulin produced by MPC11 (low producers). Only one of the MPC11 mutants (a nonproducer) showed no evidence of DNA rearrangement in or near the expressed IgH gene. The formerly expressed gamma 2b gene had been deleted in 18 of the 19 nonproducers. All of the low producers had undergone DNA rearrangement in or near the expressed IgH gene, and three of them produced immunoglobulin of a new heavy chain class. The cause for reduced heavy-chain synthesis in the low producers is not yet known. However, in several of these mutants, the defect appeared to be posttranscriptional. In these cell lines, steady-state IgH mRNA levels were much lower than in the parent cell line, while the heavy-chain gene transcription rate remained unchanged.

Identification of DJ-1/Park-7 as a Determinant of Tnfα-Induced and Stroma-Dependent Apoptosis in Leukemia Using Mass Spectometry and Phosphopetide Analysis.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1796-1796
Author(s):  
A. Mario Q. Marcondes ◽  
Li Xiang ◽  
Brian P Milless ◽  
H. Joachim Deeg
Keyword(s):  
Cell Line ◽  
Stress Responses ◽  
Time Course ◽  
Caspase 3 ◽  
Cellular Transformation ◽  
Leukemic Cells ◽  
Parent Cell ◽  
Low Grade ◽  
Marrow Stroma ◽  
Kg1 Cells

Abstract The bone marrow microenvironment provides essential signals for the fate of normal hematopoietic and for leukemic cells. Contact with marrow stroma, which is part of the microenvironment, is generally thought to convey anti-apoptotic signals to (clonal) leukemia cells. Patients with low-grade myelodysplastic syndrome (MDS) early in the disease course show high rates of apoptosis in normal and clonal marrow cells, mediated by tumor necrosis factor alpha (TNFα) and other cytokines. As MDS advances and evolves to leukemia, clonal cells tend to become apoptosis resistant. We showed previously that the leukemia-derived cell line KG1a was resistant to TNFα-mediated apoptosis, but TNFα did induce caspase-3 activation and apoptosis in KG1a cells when co-cultured with the human marrow stroma cell line HS5 (derived from healthy marrow). Apoptosis was contact dependent and required expression of TNF receptor 1 on KG1a cells. Identical results were obtained in co-cultures with primary stroma cells. Gene expression profiling of KG1a cells showed that stroma contact resulted in significant upregulation of genes involved in apoptosis, including PYCARD and p53. To further dissect the relevant signaling pathways, we used a PhosphoScan proteomic LC-MS (Liquid chromatography-mass spectrometry) method to identify proteins that were phosphorylated in response to stroma contact. In parallel to KG1a we examined the parent cell line KG1, which is sensitive to TNFα mediated apoptosis. We determined the phosphorylation sites in proteins within the leukemic cell lines using MS2 and MS3 scans. Database searches were performed with X! Tandem and Mascot and results analyzed by PeptideProphet using data from a synthetic doubly-phosphorylated peptide as control. In KG1a cells cultured without stroma support, the peptide DJ-1/Park-7 was highly phosphorylated, and expression of p53 was inhibited as indicated by decreased levels of p53 mRNA and protein. In co-culture with stroma, KG1a cells expressed higher levels of p53 protein, and levels of phosphorylated DJ-1/ Park-7 were undetectable over a time course of 30 min to 24 hours. In apoptosis-sensitive KG1 cells constitutive DJ-1/Park-7 phosphorylation (in the absence of stroma contact) was undetectable, and p53 was expressed at higher levels than in KG1a cells, consistent with the observed activation of caspase-3 and induction of apoptosis in KG1 cells. Taken together, these data suggest that phosphorylation of DJ-1/Park-7, originally identified as an oncogene product involved in cellular transformation, oxidative stress responses, and transcriptional regulation, was associated with repression of p53 and resistance to TNFα-mediated apoptosis. The relevance of DJ-1/Park-7 (and other genes identified by the PhosphoScan proteomic method) in primary MDS cells is currently being investigated at the molecular and functional levels.

scholarly journals Reexpression of p8 Contributes to Tumorigenic Properties of Pituitary Cells and Appears in a Subset of Prolactinomas in Transgenic Mice that Hypersecrete Luteinizing Hormone

2004 ◽  
Vol 18 (10) ◽  
pp. 2583-2593 ◽  
Author(s):  
Helai P. Mohammad ◽  
Darcie D. Seachrist ◽  
Christine C. Quirk ◽  
John H. Nilson
Keyword(s):  
Transgenic Mice ◽  
Cell Line ◽  
Cell Lines ◽  
Pituitary Tumors ◽  
Gh3 Cells ◽  
Parent Cell ◽  
Pituitary Cells ◽  
Wild Type ◽  
Altered Expression ◽  
Pituitary Tumorigenesis

Abstract Targeted overexpression of LH in transgenic mice causes hyperproliferation of Pit-1-positive pituitary cells and development of functional adenomas. To characterize gene expression changes associated with pituitary tumorigenesis, we performed microarray studies using Affymetrix GeneChips comparing expression profiles from pituitary tumors in LH-overexpressing mice to wild-type control pituitaries. We identified a number of candidate genes with altered expression in pituitary tumors. One of these, p8 (candidate of metastasis-1), encodes a native high-mobility group-like transcription factor previously shown to be necessary for ras-mediated transformation of mouse embryonic fibroblasts and also implicated in breast cancer progression. Herein, we show that expression of p8, normally quiescent in adult pituitary, localizes to tumor foci containing lactotropes, suggesting a linkage with their transformation. To further establish the functional significance of p8 in pituitary tumorigenesis, we constructed several clonal cell lines with reduced expression of p8 from a parent GH3 somatolactotrope cell line. These clonal derivates, along with the parent cell line, were tested for tumorigenicity by injection into athymic mice. When compared with wild-type GH3 with higher levels of p8, GH3 cells with reduced expression of p8 displayed attenuated tumor development or failed to develop tumors at all. Similar results were obtained with gonadotrope-derived cell lines that displayed reduced expression of p8. Together, these data suggest that maintenance of the transformed phenotype of pituitary GH3 cells requires expression of p8 and that it may play a similar role when reexpressed in a subset of lactotropes that form prolactinomas in vivo.

scholarly journals Signal transduction by the platelet integrin alpha IIb beta 3: induction of calcium oscillations required for protein-tyrosine phosphorylation and ligand-induced spreading of stably transfected cells.

1992 ◽  
Vol 3 (9) ◽  
pp. 989-998 ◽  
Author(s):  
A J Pelletier ◽  
S C Bodary ◽  
A D Levinson
Keyword(s):  
Signal Transduction ◽  
Molecular Weight ◽  
Cell Line ◽  
Tyrosine Phosphorylation ◽  
Calcium Oscillations ◽  
Parent Cell ◽  
Protein Tyrosine Phosphorylation ◽  
Transfected Cells ◽  
293 Cells ◽  
Stimulation Of

We demonstrate an example of signal transduction by an integrin and have begun to define the pathway through which this signaling is achieved. We constructed a stably transfected derivative of 293 cells (ATCC 1573) that expresses the platelet integrin GPIIbIIIa (alpha IIb beta 3). This cell line, clone B, adheres to and spreads on fibrinogen, a ligand for alpha IIb beta 3, while the parent cell line does not. Stimulation of these cells either by adhesion to fibrinogen or with antiserum directed against alpha IIb beta 3 results in induction of calcium oscillations, followed by tyrosine phosphorylation of at least one protein of molecular weight approximately 125 kDa. We establish that this phosphorylation, as well as the morphological rearrangements, requires the mobilization of calcium.

scholarly journals The motor neuron-like cell line NSC-34 and its parent cell line N18TG2 have glycogen that is degraded under cellular stress

2020 ◽  
Author(s):  
Brigitte Pfeiffer-Guglielmi ◽  
Ralf-Peter Jansen
Keyword(s):  
Cell Line ◽  
Energy Charge ◽  
Metabolic Stress ◽  
Neuronal Cell ◽  
Glycogen Metabolism ◽  
Active Role ◽  
Parent Cell ◽  
Normal Brain ◽  
Storage Site ◽  
Key Enzyme

AbstractBrain glycogen has a long and versatile history: Primarily regarded as an evolutionary remnant, it was then thought of as an unspecific emergency fuel store. A dynamic role for glycogen in normal brain function has been proposed later but exclusively attributed to astrocytes, its main storage site. Neuronal glycogen had long been neglected, but came into focus when sensitive technical methods allowed quantification of glycogen at low concentration range and the detection of glycogen metabolizing enzymes in cells and cell lysates. Recently, an active role of neuronal glycogen and even its contribution to neuronal survival could be demonstrated. Our studies continue these investigations on the function and regulation of neuronal glycogen metabolism. We demonstrate the presence of an active glycogen metabolism in the neuronal cell lines NSC-34 and N18TG2 and the mobilization of the glycogen stores under hypoxia, oxidative and acidic metabolic stress. The key enzyme in glycogen degradation is glycogen phosphorylase. Neurons express only the brain isoform (GPBB) that is supposed to be activated primarily by the allosteric activator AMP and less by covalent phosphorylation via the cAMP cascade. Our results indicate that neuronal glycogen is not degraded upon hormone action but by factors lowering the energy charge of the cells directly.

scholarly journals Succinate accumulation links mitochondrial MnSOD depletion to aberrant nuclear DNA methylation and altered cell fate

2020 ◽  
Author(s):  
Kimberly L. Cramer-Morales ◽  
Collin D. Heer ◽  
Kranti A. Mapuskar ◽  
Frederick E. Domann
Keyword(s):  
Cell Line ◽  
Cell Fate ◽  
Nuclear Dna ◽  
Human Cell Line ◽  
Significant Inhibition ◽  
Dna Demethylation ◽  
Parent Cell ◽  
Erythroid Precursor ◽  
Mitochondrial Superoxide ◽  
Tet Enzymes

AbstractPrevious studies showed that human cell line HEK293 lacking mitochondrial superoxide dismutase (MnSOD) exhibited decreased succinate dehydrogenase (SDH) activity, and mice lacking MnSOD displayed significant reductions in SDH and aconitase activities. Since MnSOD has significant effects on SDH activity, and succinate is a key regulator of TET enzymes needed for proper differentiation, we hypothesized that SOD2 loss would lead to succinate accumulation, inhibition of TET activity, and impaired erythroid precursor differentiation. To test this hypothesis, we genetically disrupted the SOD2 gene using the CRISPR/Cas9 genetic strategy in a human erythroleukemia cell line (HEL 92.1.7) capable of induced differentiation toward an erythroid phenotype. Cells obtained in this manner displayed significant inhibition of SDH activity and ~10-fold increases in cellular succinate levels compared to their parent cell controls. Furthermore, SOD2−/− cells exhibited significantly reduced TET enzyme activity concomitant with decreases in genomic 5-hmC and corresponding increases in 5-mC. Finally, when stimulated with δ-aminolevulonic acid (δ-ALA), SOD2−/− HEL cells failed to properly differentiate toward an erythroid phenotype, likely due to failure to complete the necessary global DNA demethylation program required for erythroid maturation. Together, our findings support the model of an SDH/succinate/TET axis and a role for succinate as a retrograde signaling molecule of mitochondrial origin that significantly perturbs nuclear epigenetic reprogramming and introduce MnSOD as a governor of the SDH/succinate/TET axis.

Sign in / Sign up

Export Citation Format

Share Document