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Product Data Sheet MCF- 7/GFP Cell Line CATALOG NUMBER: AKR-211 STORAGE: Liquid nitrogen Note: For best results begin culture of cells immediately upon receipt. If this is not possible, store at -80ºC until first culture. Store subsequent cultured cells long term in liquid nitrogen. QUANTITY & CONCENTRATION: 1 mL, 1 x 106 cells/mL in 70% DMEM, 20% FBS, 10% DMSO Background MCF-7 is a human breast cancer cell line that was first isolated in 1970 from the malignant adenocarcinoma breast tissue of a 69-year old woman. MCF-7 is the acronym of Michigan Cancer Foundation - 7, referring to the institute in Detroit where the cell line was established. MCF-7 cells are useful for in vitro breast cancer studies because the cell line has retained several ideal characteristics particular to the mammary epithelium. These include the ability for MCF-7 cells to process estrogen via estrogen receptors. MCF-7 cells are also sensitive to cytokeratin. When grown in vitro, the cell line is capable of forming domes and the epithelial like cells grow in monolayers. Growth can also be inhibited using tumor necrosis factor alpha (TNF alpha). Our MCF-7/GFP cell line stably expresses GFP; the gene was introduced using lentivirus. Figure 1. MCF-7/GFP Cell Line. Left: GFP Fluorescence; Right: Phase Contrast. Quality Control This cryovial contains at least 1.0 × 106 MCF-7/GFP cells as determined by morphology, trypan-blue dye exclusion, and viable cell count. The MCF-7/GFP cells are tested free of microbial contamination. Medium 1. Culture Medium: D-MEM (high glucose), 10% fetal bovine serum (FBS), 0.1 mM MEM NonEssential Amino Acids (NEAA), 2 mM L-glutamine, 1% Pen-Strep. 2. Freeze Medium: 70% DMEM, 20% FBS, 10% DMSO. Methods Establishing MCF-7/GFP Cultures from Frozen Cells 1. Place 10 mL of complete DMEM growth medium in a 50-mL conical tube. Thaw the frozen cryovial of cells within 1–2 minutes by gentle agitation in a 37°C water bath. Decontaminate the cryovial by wiping the surface of the vial with 70% (v/v) ethanol. 2. Transfer the thawed cell suspension to the conical tube containing 10 ml of growth medium. 3. Collect the cells by centrifugation at 1000 rpm for 5 minutes at room temperature. Remove the growth medium by aspiration. 4. Resuspend the cells in the conical tube in 15 mL of fresh growth medium by gently pipetting up and down. 5. Transfer the 15 mL of cell suspension to a T-75 tissue culture flask. Place the cells in a 37°C incubator at 5% CO2. 6. Monitor cell density daily. Cells should be passaged when the culture reaches 95% confluence. Recent Product Citations 1. Singh, A. P. et al. (2016). Quantitative characterization of in vitro bystander effect of antibodydrug conjugates. J Pharmacokinet Pharmacodyn. doi:10.1007/s10928-016-9495-8. 2. Ansari, A. et al. (2016). A method of targeted cell isolation via glass surface functionalization. J Vis Exp. doi:10.3791/54315. 3. Nwokeoha, S. et al. (2016). The application of clinical lithotripter shock waves to RNA nucleotide delivery to cells. Ultrasound Med Biol. doi:10.1016/j.ultrasmedbio.2016.06.001. 4. Ambrose, J. et al. (2015). Mediated coalescence: a possible mechanism for tumor cellular heterogeneity. Am J Cancer Res. 5:3485-3504. 5. Shabo, I. et al. (2015). Macrophage traits in cancer cells are induced by macrophage-cancer cell fusion and cannot be explained by cellular interaction. BMC Cancer. 15:922. 6. Spencer, A. et al. (2015). A high-throughput mechanofluidic screening platform for investigating tumor cell adhesion during metastasis. Lab Chip. doi:10.1039/C5LC00994D. 7. Larson, B. & Sherman, H. (2015). Automated, Kinetic Imaging of Cell Migration and Invasion Assays using Corning® FluoroBlok™ Inserts. Corning Application Note. 8. Ansari, A. et al. (2015). Secondary anchor targeted cell release. Biotechnol Bioeng. doi:10.1002/bit.25648. 9. Zhang, K. et al. (2014). Block-cell-printing for live single-cell printing. PNAS. 111:2948-2953. 10. Zhang, W. et al. (2012).Microfluidics separation reveals the stem-cell–like deformability of tumorinitiating cells. PNAS. 109:18707-18712. Warranty These products are warranted to perform as described in their labeling and in Cell Biolabs literature when used in accordance with their instructions. THERE ARE NO WARRANTIES THAT EXTEND BEYOND THIS EXPRESSED WARRANTY AND CELL BIOLABS DISCLAIMS ANY IMPLIED WARRANTY OF MERCHANTABILITY OR WARRANTY OF FITNESS FOR PARTICULAR PURPOSE. CELL BIOLABS’s sole obligation and purchaser’s exclusive remedy for breach of this warranty shall be, at the option of CELL BIOLABS, to repair or replace the products. In no event shall CELL BIOLABS be liable for any proximate, incidental or consequential damages in connection with the products. This product is for RESEARCH USE ONLY; not for use in diagnostic procedures. Contact Information Cell Biolabs, Inc. 7758 Arjons Drive San Diego, CA 92126 Worldwide: +1 858-271-6500 USA Toll-Free: 1-888-CBL-0505 E-mail: [email protected] www.cellbiolabs.com 2010-2016: Cell Biolabs, Inc. - All rights reserved. No part of these works may be reproduced in any form without permissions in writing.