Figure 143.

Single cell preparations from human tumor vs. non-tumor tissues and characterisation of human tumor vs. non-tumor epithelial cells. (A) Human tumor (upper row) and adjacent non-tumor tissue (lower row) was obtained as surplus tissue in the course of a pulmonary tumor resection with informed consent (MHH Nr. 1747). After tissue digestion, single cells were stained with a live/dead dye (QDot585) and anti-human CD45 (Alexa-Fluor700) mAb. The hierarchical gating strategy starts with exclusion of doubles and aggregates in the FSC-A/FSC-H plot, followed by exclusion of dead cells in the QDot585/SSC-A plot and leukocytes, i.e., CD45-positive cells in the CD45/SSC-A plot. The remaining living CD45-negative single cells are shown in the FSC-A/SSC-A plot and in the blue gate, epithelial cells including tumor cells in the tumor tissue, can be identified according to their relative size and granularity. (B) A renal tubular cancer cell (RTCC) and the corresponding non-tumor tubular cell line (RNTC) derived from tumor and adjacent non-tumor tissue of the same patient are compared with respect to surface expression of the following markers: HLA class I (mAb W6/32) and the adhesion molecule ICAM-1 (CD54, mAb gp89). All primary mAb are mouse IgG2a and were stained with a goat-anti-mouse PE-labeled secondary Ab.