McCoy cells

Substances capable of interaction with ROS are claimed to play an important role in the prevention of cancer. Some compoimds of the unsaponifiable fraction from virgin oil (oleuropein, tyrosol, sterols and triterpenoids) were assayed to determine their cytostatic activity in McCoy cells (originated from synovial fluid from a patient suffering degenerative arthritis) [92]. Oleuropein and the sterol+triterpenoid fraction potently inhibited cell growth at 6 Xg/ml (83 and 89% respectively), a concentration recommended by the protocols of the National Cancer Institute of USA [93]. IC50 for oleuropein and sterol+terpenoid fraction was 4.4 and 0.11 ig/ml, respectively. Tyrosol exhibited lower inhibition IC50 was 10 pg/ml. 

Although tyrosol does not surpass the antioxidant capacity of hydroxytyrosol, it exhibits cytostatic activity against McCoy cells and potential activity as an anti-tumoral compound [237]. Also, it exerts antiarrhythmic [238] and cardioprotective actions by preventing stressor-induced change at the level of cyclic nucleotides in the myocardium [239]. 

SK-BR-3 This is an adherent breast carcinoma cell line. This cell line is cultured in McCoy s 5A medium with 2mM glutamine and 10% Fetal Bovine Serum (FBS) at 37°C in a 5% C02 in air atmosphere. 

Singh, N.P., McCoy, M.T., Tice, R.R. and Schneider, E.L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research 175 184-191. 

HCT-116 human colon carcinoma (ATCC, Bethesda MD) cells were grown in McCoy s 5A) and were routinely subcultured twice weekly. Antiproliferative assay was performed by chemoluminescence assay based on quantification of ATP. Cells in their exponential phase of growth were treated at different times (lh or 24h) with different concentrations of edotecarin or SN-38. For post-treatment recovery studies, cells were washed with PBS and left in drug-free culture medium. Then, cell medium was collected to avoid any cell loss. Cells in monolayer were washed, detached with trypsin, and collected in the medium. Cells were counted in a Multisizer 3 Coulter Counter to measure the drug s effects on growth inhibition. Samples were fixed either... 

In K562 cells TPA activated the MDRl promoter and led to increased MDRl mRNA levels. This activation was mediated by the zinc finger transcription factor EGRl. This activation by TPA was inhibited by the Wilms Tumor suppressor WTl (McCoy et al., 1995 McCoy et al., 1999). Controversial response to TPA may depend on the presence or absence of EGRl or WTl in the cell lines investigated. 

McConkey DJ, Hartzell P, Jondal M, Orrenius S (1989) Inhibition of DNA fragmentation in thymocytes and isolated thymocyte nuclei by agents that simulate protein kinase C.) Biol Chem 264 13399-13402 McConkey DJ, Nicotera P, Orrenius S (1994) Signalling and chromatin fragmentation in thymocyte apoptosis. Immunol Rev 142 343-363 McCoy C, Smith DE, Cornwell MM (1995) 12-0-tetradecanoylphorbol-13-acetate activation of the MDRl promoter is mediated by EGRl. Mol Cell Biol 15 6100-6108... 

Cell culture medium McCoy s 5A Medium with L-glutamine, 10% FCS. 

Aspirate the supernatant and resuspend the cell pellet in freezing medium (McCoy s 5A Medium with L-glutamine, 10% PCS, 5% DMSO) to a concentration of 1 x 10 cells/ml. 

HCT116 cells and culture media (McCoy s+ 2 mM L-glutamine+ 10% PCS). 

RPMI, modified McCoy 5 A medium for cell culture HEPES, 4-(2-hydroxyethyl)piperazine-l-ethanesulfonic acid IMDM, Iscov s modified Dulbecco s medium PCS, fetal calf serum... 

R. D. McCoy, E. R. Vimr, and F. A. Troy, CMP-NeuNAc poly-a-2,8-sialosyl sialyltransferase and the biosynthesis of poiysialosyl units in neural cell adhesion molecules, J. Biol. Chem. 260 i 2695 (1985). 

Shelburne CP, McCoy ME, Piekorz R, Sexl W, et al. 2002. STAT5 An essential regulatory of mast cell biology. Mol Immunol. 38 1187-1191. 

Lu, Z., Murray, K. S., Van Cleave, V., LaVallie, E. R., Stahl, M. L., and McCoy, J. M. (1995) Expression of thioredoxin random peptide libraries on the Escherichia coli cell surface as functional fusions to flagellin a system designed for exploring protein-protein interactions. Biotechnology 13, 366-372. 

Grow the HT-29 cell line in T75 flasks in McCoy s 5A medium (10% FBS). When cells are confluent, harvest HT-29 cells by adding 2.0 ml of TrypLE Express. After the cells detach from the flasks, resuspend the cells in McCoy s medium (10% FBS) to inactivate the TrypLE Express. Count the cell concentration as described below. Prepare two small T25 flasks with McCoy s medium (10% FBS) and seed 1 x 106 cells per flask. Keep the HT-29 cells in culture as they will be needed again later. 

Aspirate HT-29LP medium (McCoy s 5A, supplemented with 1% glutaMAX, 10% FBS, and 0.8 mg/ml geneticin), from adherent HT-29LP cells growing in a polystyrene flask or dish and add 2 ml TrypLE Express. Move the solution around the plate and quickly aspirate. 

Add medium (McCoy s 5A, supplemented with 1% Gluta-Max and 10% FBS without geneticin) to bring HT-29LP cells to the final desired concentration. For the utilized dose of 1 x 106 cells/mouse, the cells should be brought to 20 x 106 cells/ml. Put the cells on ice to bring to approximately 4°C. 

Fraser, J. H., Rincon, M., McCoy, K. D., and Le Gros, G. 1999. CTLA4 ligation attenuates AP-1, NFAT and NF-kappaB activity in activated T cells. Eur J Immunol 29 838-844. 

Thus McCoy s medium 5A (McCoy et al., 1959) has been used as a standard medium for cloning cells. It is based on BME amino acids and the vitamins from medium 199 (Appendix 1 Table 18). This was modified further to form RPMI (Rosswell Park Memorial Institute)-1629 (Appendix 1 Table 17) for long-term culture of leukaemic myoblasts (Armstrong, 1966). 

Exponentially growing cells are used and it is important to maintain constant pH and temperature throughout the selection procedure. Klevecz et al. (1974, 1975) have used CHO cells growing in McCoy s medium with 20% foetal bovine serum and Hepes buffer. They maintain the cells throughout in this medium at 37°C and, of the cells they select, 98-99% are in mitosis. The viability of these selected cells approaches 100% and on reseeding half the cells attach within 1 h of selection and maximum attachment is found by 4 h (Klevecz, 1975). However, they select a very small proportion of the original cells. 

Inoculate cells into bottles 24 h prior to initial selection. 5 X 107 Don-C Chinese hamster fibroblast cells may be inoculated into a roller bottle in McCoy s medium (see 11.2.2). 

Subculture Don-C cells every 24 h in McCoy s 5a medium (Appendix 1) containing 20% foetal calf serum and 0.08% lactalbumin hydrolysate, seeding cells at 1.2 X 105/ml(3 X 104 cells/cm2). 

Bystander activation activation of autoreactive cells through nonspecific inflammation and induction of inflammatory cytokines and chemokines is also of pathological consequence in MS. It has been suggested that bystander activation, induced by persistent virus infection or primed by molecular mimicry may activate autoreactive T-cells specific for the CNS (McCoy et al., 2006). Einally, cryptic antigens may also play a role in immune activation. In other immune-mediated diseases such as Chronic Lymphocytic Thyroiditis and Chagas Heart Disease, exposure of cryptic epitopes leads to the activation of autoimmune cells and further contributes to... 

Pollice, A A, McCoy, J. P, Jr., Shackney, S E, Smith, C A., Agarwal, J., Burholt, D R., Janocko, L. E, Hornicek, F J, Singh, S. G, and Hartsock, R. J. (1992) Sequential paraformaldehyde and methanol fixation for simultaneous flow cytometric analysis of DNA, cell-surface proteins, and intracellular proteins. Cytometry 13,432-444... 

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