Pe labeled mouse anti human cd29

Flow cytometry was applied to compare the expression of a panel of CSC markers, some of which are known renal CSC markers, including CD24, CD44, CD133, CD105, CXCR4, and CD73 (23 (link)). Other cell surface markers such as CD29, CD34, CD56 (NCAM), CD90, CD117, and CD146 have been found to be differentially expressed between PCs and SDCs of other cancers (24 (link)). The SDCs were dissociated by accutase and then washed with PBS. Nearly 1 × 10⁵ single viable cells from each population were incubated with 1 μg/ml fluorescently labeled monoclonal antibodies, or respective isotype controls, in the dark for 30 min on ice. Several antibodies were used: phycoerythrin (PE)-labeled mouse anti-human CD24, PE-labeled mouse anti-human CD29, FITC-labeled mouse anti-human CD34, fluorescein iso thiocyanate (FITC)-labeled mouse anti-human CD44, PE-labeled mouse anti-human CD56, PE-labeled mouse anti-human CD73, FITC-labeled mouse anti-human CD90, PE-labeled mouse anti-human CD105, PE-labeled mouse anti-human CD117, PE-labeled mouse anti-human CD133, PE-labeled mouse anti-human CD146, and PE-labeled mouse anti-human CXCR4 (all, BD Biosciences, USA, except CD90, DAKO, Denmark). Then, the cells were washed, resuspended, and analyzed by flow cytometry (FACS Calibur, BD, USA). The FlowJo Version 7 was also used to analyze the data.

Flow cytometry was used to compare the protein expression of a panel of suggested renal CSCs and differentially expressed markers between SDCs and their PCs in other cancers (20 (link)). The following fluorophore-conjugated mouse antihuman monoclonal antibodies were used for flow cytometry: phycoerythrin (PE)-labeled mouse antihuman CD24, fluorescein isothiocyanate (FITC)-labeled mouse antihuman CD44, PE-labeled mouse antihuman CD133, PE-labeled mouse antihuman CD105, PE-labeled mouse antihuman CXCR4, PE-labeled mouse antihuman CD56, FITC-labeled mouse antihuman CD90, PE-labeled mouse antihuman CD29, PE-labeled mouse antihuman CD73, FITC-labeled mouse antihuman CD34, PE-labeled mouse antihuman CD146 and PE-labeled mouse antihuman CD117 (All antibodies were from BD Biosciences, USA, except CD90 from DAKO, Denmark).
Also, 1×10⁵ single viable cells from live cells of PCs and SDCs were dissociated with accutase (Sigma-Aldrich, St. Louis, MO, USA) washed once with PBS, and stained with conjugated antibodies, or respective isotype controls according to the manufacturers’ recommendations. Then, the cells were washed, re suspended, and analyzed with FACSCalibur (Becton Dickinson, San Jose, CA). The FlowJo Version 7 was also used for data analysis.