Time-lapse Imaging of Primary Preneoplastic Mammary Epithelial Cells Derived from Genetically Engineered Mouse Models of Breast Cancer |
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Authors: | Rebecca E Nakles Sarah L Millman M Carla Cabrera Peter Johnson Susette Mueller Philipp S Hoppe Timm Schroeder Priscilla A Furth |
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Institution: | Department of Oncology, Georgetown University;Lombardi Comprehensive Cancer Center, Georgetown University;Stem Cell Dynamics, Helmholtz Zentrum München - German Research Center for Environmental Health;Department of Medicine, Georgetown University;Department of Nanobiomedical Science and WCU Research Center of Nanobiomedical Science, Dankook University |
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Abstract: | Time-lapse imaging can be used to compare behavior of cultured primary preneoplastic mammary epithelial cells derived from different genetically engineered mouse models of breast cancer. For example, time between cell divisions (cell lifetimes), apoptotic cell numbers, evolution of morphological changes, and mechanism of colony formation can be quantified and compared in cells carrying specific genetic lesions. Primary mammary epithelial cell cultures are generated from mammary glands without palpable tumor. Glands are carefully resected with clear separation from adjacent muscle, lymph nodes are removed, and single-cell suspensions of enriched mammary epithelial cells are generated by mincing mammary tissue followed by enzymatic dissociation and filtration. Single-cell suspensions are plated and placed directly under a microscope within an incubator chamber for live-cell imaging. Sixteen 650 μm x 700 μm fields in a 4x4 configuration from each well of a 6-well plate are imaged every 15 min for 5 days. Time-lapse images are examined directly to measure cellular behaviors that can include mechanism and frequency of cell colony formation within the first 24 hr of plating the cells (aggregation versus cell proliferation), incidence of apoptosis, and phasing of morphological changes. Single-cell tracking is used to generate cell fate maps for measurement of individual cell lifetimes and investigation of cell division patterns. Quantitative data are statistically analyzed to assess for significant differences in behavior correlated with specific genetic lesions. |
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Keywords: | Cancer Biology Issue 72 Medicine Cellular Biology Molecular Biology Anatomy Physiology Oncology Mammary Glands Animal Epithelial Cells Mice Genetically Modified Primary Cell Culture Time-Lapse Imaging Early Detection of Cancer Models Genetic primary cell culture preneoplastic mammary epithelial cells genetically engineered mice time-lapse imaging BRCA1 animal model |
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