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  • Cell separation and gene expression analysis in a tumor-stroma interaction model
    Dahse, Regine ; Kosmehl, Hartwig
    A novel technique for co-culturing and separating fibroblasts and carcinoma cells in a 2-D model of tumor stroma interaction is presented. The methodology is based on cell co-cultivation on an 1.35 ... microm thin membrane followed by rapid immunostaining and microdissection of the different cell compartments using a laser microdissection system (P.A.L.M. Microlaser Technologies AG, Germany). For identifying the tumor cell compartment, immunolabeling for a marker that is expressed only in epithelial tumor cells is performed. The RNA quality from the microdissected co-cultured cells was successfully proved by RT-PCR for a housekeeping gene transcript and for the laminin gamma 2 chain gene transcript used before in the tumor cell immunostaining. Laminin cDNA wasamplificable only in tumor cells and not in the co-cultivated fibroblasts indicating no cell-cross-contamination during microdissection. Microdissected tumor and stroma cells from the presented membrane based co-culture model can be used for gene expression profiling and DNA based analysis in the investigation of tumor-stroma interactions.
    Vir: Image analysis & stereology : official journal of the International Society for Stereology. - ISSN 1580-3139 (Vol. 23, no. 3, nov. 2004, str. 153-157)
    Vrsta gradiva - članek, sestavni del
    Leto - 2004
    Jezik - angleški
    COBISS.SI-ID - 20711897

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    Digitalna knjižnica Slovenije - dLib.si

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