The CWR22Pc prostate cancer cell line was kindly provided by Marja Nevalainen (Dagvadorj et al., 2008 (link)). We found that this cell line contained a subpopulation of cells with fibroblast-like morphology that were human EpCAM-negative and confirmed to be of mouse origin. In order to purify tumor cells and mouse fibroblasts, we plated CWR22Pc at 400–800 cells per well (6-well) in 50% conditioned media. Numerous multi-clonal, cancer epithelial islands visually free of fibroblasts were isolated by cloning cylinders and then pooled to derive the pure epithelial subline, CWR22Pc-EP, in short 22Pc-EP. Human EpCAM-negative cancer-associated mouse fibroblasts were obtained by performing mouse specific H-2Kb and H-2Db MHC class I sorting (Biolegend #114608) and the FACs purified cancer-associated fibroblasts were termed as CWR22Pc-CAF, in short 22Pc-CAF. Purified 22Pc-EP and 22Pc-CAF cells were transduced with eGFP (SGEP-Renilla) (Fellmann et al., 2013 (link)) or tdTomato (QCXIp-tdTomato, Clontech #9136–1). tdTomato was derived from vector p-tdTomato (Clontech #632531) and cloned into the AgeI and EcoRI sites of QCXIP retroviral vector. Both were and selected with 1 μg/mL puromycin (Gibco #A1113803) for 5 days.
H 2db mhc class 1 sorting
Manufactured by BioLegend
H-2Db MHC class I sorting is a lab equipment used for the detection and isolation of H-2Db MHC class I molecules. It allows for the identification and separation of cells expressing this specific protein.
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Lab products found in correlation
2 protocols using h 2db mhc class 1 sorting
1
Isolation and Purification of Prostate Cancer Cell Lines
2
Isolation and Purification of Prostate Cancer Cell Lines
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The CWR22Pc prostate cancer cell line was kindly provided by Marja Nevalainen (Dagvadorj et al., 2008 (link)). We found that this cell line contained a subpopulation of cells with fibroblast-like morphology that were human EpCAM-negative and confirmed to be of mouse origin. In order to purify tumor cells and mouse fibroblasts, we plated CWR22Pc at 400–800 cells per well (6-well) in 50% conditioned media. Numerous multi-clonal, cancer epithelial islands visually free of fibroblasts were isolated by cloning cylinders and then pooled to derive the pure epithelial subline, CWR22Pc-EP, in short 22Pc-EP. Human EpCAM-negative cancer-associated mouse fibroblasts were obtained by performing mouse specific H-2Kb and H-2Db MHC class I sorting (Biolegend #114608) and the FACs purified cancer-associated fibroblasts were termed as CWR22Pc-CAF, in short 22Pc-CAF. Purified 22Pc-EP and 22Pc-CAF cells were transduced with eGFP (SGEP-Renilla) (Fellmann et al., 2013 (link)) or tdTomato (QCXIp-tdTomato, Clontech #9136–1). tdTomato was derived from vector p-tdTomato (Clontech #632531) and cloned into the AgeI and EcoRI sites of QCXIP retroviral vector. Both were and selected with 1 μg/mL puromycin (Gibco #A1113803) for 5 days.
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