Embryonic fibroblast cells

Reconstitution of full-length BRCAl into mouse embryonic fibroblast cells with a disrupted BRCAl led to an increase in resistance to several DNA damaging agents, including the platinum compounds carboplatin and oxaliplatin, the topoisomerase 1 drugs irinotecan and topotecan, and the topoisomerase 11 drugs doxorubicin and etoposide (61). 

Irradiated embryonic fibroblast feeder cell layer to be used for growing the embryonic stem cell line G418 resistant embryonic fibroblast cells are derived from embryos that carry a targeted disruption such as the 3-2 microglobulin gene (9). 

Approximately 105 transfected embryonic stem cells are selected. Transfected embryonic stem cells are grown on layers of y-irradiated embryonic fibroblast cells. Transfected cells are plated on G-418 resistant embryonic fibroblasts feeder layers and selected in G-418 and after 36 h they are selected in Fialuridine (FIAU) or gancyclovir. 

Naim, R. lida, T. Takahashi, A. Honda, T. Monoda-nsylcadaverine inhibits cytotoxicity of Vibrio parahaemo-lytiaiStbstmosX > c direct hemolysin on cultured rat embryonic fibroblast cells. FEMS Microbiol. Lett. 2001,196, 99-105. 

Imidazole antimycotics, ketoconazole, clotrimazole, and miconazole are potent inhibitors of various cytochrome P450-isoenzymes that also affect the metabolism of retinoids. They were fust shown to inhibit the metabolism of RA in F9 embryonal carcinoma cells. When tested in vitm liarazole, a potent CYP-inhibitor, suppressed neoplastic transformation and upregulated gap junctional communication in murine and human fibroblasts, which appeared to be due to the presence of retinoids in the serum component of the cell culture medium. Furthermore, liarazole magnified the cancer chemopreventive activity of RA and (3-carotene in these experiments by inhibiting RA-catabolism as demonstrated by absence of a decrease in RA-levels in the culture medium in the presence of liarazole over 48 h, whereas without liarazole 99% of RA was catabolized. In vivo, treatment with liarazole and ketoconazole reduced the accelerated catabolism of retinoids and increased the mean plasma all-irans-RA-concentration in patients with acute promyelocytic leukemia and other cancels. 

The biological impact of starch capped copper nanoparticles on mouse embryonic fibroblast (3T3L1) cells in vitro) was also evaluated by various parameters. More than 85 % of the 3T3Llcells were found to be viable, even after 20 hours time exposure which implies minimum impact on cell viability and morphology. The study demonstrates dose dependent cytotoxic potential of SCuNPs, that is non cytotoxic in the nanogram dose and moderately cytotoxic in the microgram doses (Fig. 10). Comparison of SCuNPs with Cu ions and uncapped copper nanoparticles (UCuNPs) revealed that, ions are more cytotoxic than SCuNPs. This observation supports the theory of slow release of ions from starch coated nanoparticles. 

Fig. 10. Effect of starch capped copper nanoparticles on cell viability (MTT assay) in case of mouse embryonic fibroblast (3T3L1).

Fig. 11. Photomicrographs of mouse embryonic fibroblast (3T3L1) cells (A) untreated, treated with (B) control (C) SOOng CuNPs, (D) lOOOng CuNPs.

It is well known that 1,10-phenanthrolines are highly active ironchelating agents. The parent compound itself has recently been shown to increase HIF-la levels in ocular tissue and to suppress 02-mediated epithelial cell proliferation when administered to mice [29]. A quantitative assay was developed to measure transcriptional potency of certain HIF stabilizers via an HRE-mediated (3-lactamase production in which the EC50 of 1,10-phenanthroline was measured to be approximately 8 pM. In addition, VEGF was dose-dependently produced in mouse embryonic fibroblasts by 1,10-phenanthroline with an EC50 of... 

Figure 14.19 Simplified schematic overview of the directed differentiation of human embryonic stem cells to form differentiated dopamine like neurons. The full pathway details are available in Yan, Y., Yang, D., Zarnowska, E.D., Du, Z., Werbel, B., Valliere, C., etai 2005. Directed differentiation of dopaminergic neuronal subtypes from human embryonic stem cells. Stem Cells 23, 781-790. FGF fibroblast growth factor SHH sonic hedgehog (a regulatory protein)...

Immortalized embryonic fibroblasts from PTEN-/- mice were shown to have elevated PtdIns(3,4,5)P3 level, constitutively elevated PKB/AKT activity and a decreased sensitivity to cell death in response to apoptotic stimuli (Stambolic et al, 1998). Homozygotic mutation of PTEN induced... 

In vitro studies with unmodified and modified N-terminal peptides of H3 demonstrated that Lys-14 acetylation did not interfere with methylation at Lys-9 by Suv39hl, while phosphorylation at Ser-10 and acetylation at Lys-9 did (Fig. 7). Further dimethylation of Lys-9 reduced enzymatic activity [186], A Suv39h double null primary mouse embryonic fibroblasts had higher levels of Ser-10 phos-phorylated H3 than wild type cells. These mutant cells had increased numbers of micro- and polynuclei. Oversized nuclei were characteristic of subpopulation of cells. The level of Lys-9 methylated FI3 in wild type cells and Suv39h double null cells was similar, demonstrating that other FI3 methyltransferases were involved [195]. Phosphorylation of Ser-10 by Ipll/aurora was also studied. Acetylation at Lys-14 promoted the activity of the mitotic kinase, while dimethylation, but not acetylation at Lys-9, reduced activity of the kinase [186]. 

Irradiated wild type (WT) and CD 154 expressing (CD 154) NIH3T3 mouse embryonic fibroblasts (see Note 2) WT and CD154 NIH3T3 cells are cultured in complete DMEM and maintained in a humidified atmosphere of 5% CO at 37°C. 

Feeder cells G418 resistant murine embryonic fibroblasts. 

Kosbash It is fly behaviour. If we did Paolo Sassone-Corsi s experiment and took out the Drosophila embryonic fibroblasts, we could study individual cell function. 

In the EST, two murine cell lines are used to assess teratogenic potential the embryonic D3 stem cell (ES) which represents the embryonic tissue and the 3T3 fibroblast cell which represents the adult tissue. The D3 cells are maintained in an undifferentiated stage in the presence of leukemia inhibiting factor (LIE), then released from LIE and allowed to form embryo bodies that differentiate into cardiomyocytes. The D3 and 3T3 cells are exposed to a range of concentrations of the test ingredient. After a 10-day... 

Spielmann H, Pohl I, Doering B, Liebsch M, Moldenhauer E (1997) The embryonic stem cell test, an in vitro embryotoxicity test using two permanent mouse cell lines, 3 T3 fibroblasts and embryonic stem cells. In Vitro Toxicol 10 119-127... 

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