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Cell separation and gene expression analysis in a tumor-stroma interaction modelDahse, Regine ; Kosmehl, HartwigA 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 (Letn. 23, št. 3, 2004, str. 153-157)Vrsta gradiva - članek, sestavni delLeto - 2004Jezik - angleškiCOBISS.SI-ID - 20711897
Avtor
Dahse, Regine |
Kosmehl, Hartwig
Teme
Coculture |
Fibroblasts |
Tumor Cells, Cultured |
Neoplasm Invasiveness |
Gene Expression |
Immunohistochemistry |
Microscopy, Confocal |
Fibroblasti |
Gensko izražanje |
Kokultura |
Novotvorba, invazivnost |
Tumorske celične kulture |
Imunohistokemija |
Mikroskopija konfokalna
vir: Image analysis & stereology : official journal of the International Society for Stereology. - ISSN 1580-3139 (Letn. 23, št. 3, 2004, str. 153-157)
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