19 KB356 KB15 KB200020242004Dahse, RegineKosmehl, Hartwigletnik:23številka:35 stranistr. 153-157ISSN:1580-3139COBISSID:20711897URN:URN:NBN:SI:doc-80ENNJJ5enDruštvo za stereologijo in kvantitativno analizo slike, Medicinska fakultetaImage analysis and stereologyCocultureFibroblastiFibroblastsGene ExpressionGensko izražanjeImmunohistochemistryImunohistokemijaizražanje genovKokulturalaserska mikrodisekcijaMicroscopy, ConfocalMikroskopija konfokalnaNeoplasm InvasivenessNovotvorba, invazivnostTumor Cells, CulturedtumorjiTumorske celične kultureCell separation and gene expression analysis in a tumor-stroma interaction model|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 interactionsTEXTznanstveno časopisjejournalsSlovenian National E-content AggregatorNational and University Library of SloveniaDruštvo za stereologijo in kvantitativno analizo slike